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Asymetrix ToolBook File | 1994-06-09 | 431KB | 5,981 lines

INITIALISE 4protease,substrate1,substrate2,promoter,tadd,planinit GET SETTINGS FROM DIALOG BOX dlginit retvalue dialog(dlgbox MODIFY getvalue( BOK") Bstraina") /"fermenter" "(A)" "MATa, ura3" Bstrainb") "(B)" q, pep4, prb1" Bstrainc") "(C)" Badh") "ADH2" Bgal") "GAL1" Bpgk") "PGK1" Bethanol") "Ethanol" Bgalactose") "Galactose" Bglucose") "Glucose" Bethanol2") Bgalactose2") Bglucose2") ,"combobox --- "&"& "2 hours"& k&"4 k&"6 k&"8 k&"10 k&"12 k&"14 k&"16 k&"18 k&"20 k&"22 k&"24 k&"26 k&"28 k&"30 k&"32 k&"34 k&"36 k&"38 k&"40 setvalue( ",x) buttonup buttonup dlginit dlgbox dialog button OK getvalue button straina getvalue protease fermenter protease fermenter MATa, ura3 button strainb getvalue ffffff protease fermenter protease fermenter MATa, ura3, pep4, prb1 button strainc getvalue 333333 protease fermenter protease fermenter MATa, ura3, pep4 button adh getvalue promoter fermenter button gal getvalue promoter fermenter button pgk getvalue promoter fermenter button ethanol getvalue substrate1 fermenter Ethanol button galactose getvalue substrate1 fermenter Galactose button glucose getvalue substrate1 fermenter Glucose button ethanol2 getvalue substrate2 fermenter Ethanol button galactose2 getvalue substrate2 fermenter Galactose button glucose2 getvalue substrate2 fermenter Glucose combobox tadd getvalue fermenter 2 hours 4 hours 6 hours 8 hours 10 hours 12 hours 14 hours 16 hours 18 hours 20 hours 22 hours 24 hours 26 hours 28 hours 30 hours 32 hours 34 hours 36 hours 38 hours 40 hours combobox tadd setvalue retvalue protease substrate1 substrate2 promoter planinit startpage MCBcoursewaree courseware MCBBBBBBeware Department of Molecular and Cell Biology University of Aberdeennnnnnnn YEAST FOREIGN GENE EXPRESSION SYSTEMS continue "letterpage" buttonUp buttonUp letterpage Start tutorial ExitProgram buttonUp buttonUp Button Zhgh, newhgh 4mouth /"labbk "one" /" >5 "expdetails" /" + 1 Zhgh >=200 "congratulations" cn<20 "bowtie" >150 "okay" <=150 "poor" 120,87,50 flipvertical enterPage leavePage enterPage labbk background labbk background expdetails labbk background labbk background congratulations congratulations bowtie 240,50,100 bowtie 60,50,100 bowtie 240,50,100 bowtie mouth flipvertical mouth newhgh leavePage congratulations mouth flipvertical mouth message so the sysBook gets txClassName c"thumb" sliderPosition "graph1" "graph2" "etoh1" "biomass" "etoh2" >0.4 "graph3" >0.5 "graph4" >0.6 "graph5" >0.7 "graph6" >0.8 >0.5 " & x) >0.4 >0.5 >0.6 >0.7 >0.8 stilldown buttonstilldown buttonUp buttonstilldown txClassName thumb sliderPosition graph1 graph2 etoh1 biomass etoh2 graph2 etoh2 graph1 graph3 etoh1 biomass 333333 graph3 biomass graph2 graph4 etoh2 333333 ffffff graph4 biomass graph3 graph5 ffffff graph5 biomass graph4 graph6 graph6 biomass graph5 etoh1 etoh2 buttonUp graph etoh1 etoh2 txClassName thumb sliderPosition graph1 etoh1 graph2 etoh2 333333 graph3 333333 ffffff graph4 ffffff graph5 graph6 labbk background reportbackground "jeanspeak" "ob4hint" 4objective1, objective2, objective3, objective4, usedbefore F>=1 B>=1 >>=1) B"ob4button" "You must read these objectives correct order. Go click on ouseEnter mouseLeave mouseEnter buttonUp mouseEnter jeanspeak ob4hint mouseLeave ob4hint buttonUp buttonUp ob4button You must read these objectives in the correct order. Go back and click on the first objective. You must read these objectives in the correct order. Go back and click on the first objective. You must read these objectives in the correct order. Go back and click on the first objective. You must read these objectives in the correct order. Go back and click on the second objective. You must read these objectives in the correct order. Go back and click on the third objective. objective1 objective2 objective3 objective4 usedbefore 4inoc,checkgraph,findpage,nosave "medium" "Do you really want inoculate a dirty, empty fermenter?" f"No" eatclicks() "autoclave tape" "Your culture has turned a strange colour because forgot sterilise the O'd better begin f"Begin "labtxt" increases H1 each drawn data points (rectangles) called "newgraph" "arrow" store results experiment your f"Yes" --count graphs saved labbook /"labbk JPage H2000,-250 "checkprint" "studname" "expdetails" " " & "promoter" " " & "protease" " " & "substrate1" " " & "substrate2" " " & "tadd" " " & "yield" "run1" " " & "ethanol" " " & buttonUp buttonUp medium Do you really want to inoculate a dirty, empty fermenter? eatclicks autoclave tape medium 120,50,100 Your culture has turned a strange colour because you forgot to sterilise the fermenter and medium! You'd better begin again. Begin again! eatclicks labtxt buttonUp inoculate fermenter arrow group newgraph Do you want to store the results of this experiment in your lab book? paste labbk background newgraph labbk background labbk background checkprint studname checkprint studname checkprint studname labbk background paste background expdetails labbk background paste expdetails labbk background promoter fermenter expdetails labbk background protease fermenter expdetails labbk background substrate1 fermenter expdetails labbk background substrate2 fermenter expdetails labbk background fermenter expdetails labbk background yield expdetails labbk background ethanol expdetails labbk background checkprint studname medium fermenter autoclave tape fermenter eatclicks checkgraph findpage nosave ExitProgram buttonUp buttonUp "helppage" buttonUp buttonUp helppage "aims" buttonUp buttonUp buttonUp buttonUp run button Laboratory buttonUp buttonUp background background button buttonUp buttonUp buttonUp background button Library q button "questions1" buttonUp buttonUp questions1 Report buttonUp buttonUp buttonUp q button "results1" buttonUp buttonUp results1 results button Results buttonUp buttonUp buttonUp q button buttonUp buttonUp buttonUp q button --- DECLARE SYSTEM VARIABLES --- Enter fthese --- Substrate: 10=Glc 7=Gal 5=EtOH --- substrate1= ; substrate2= " values also serve %conversion factors calculating --- lags, growth rates v utilisation --- Promoter: 10=PGK 7=GAL 5=ADH --- tadd= addition , must be a multiple --- protease: 0.65=high 0.85=medium 0.95=low ,promoter, E,culturecolor AND SET LOCAL Zable,t,u,p,x,glc,gal,etoh,s, ,lag,umax, --- Set I(t), (u), product concn (p), biomass (x), --- glucose j), galactose w), ethanol Ps), identity ( --- ( calculated main loop) --- Move arrow 2500+ *80,1000 --- Clear out yield "0 units" "0 mg/ml" --- MAIN LOOP --- Repeat --- SUBSTRATE CONCENTRATION --- Preference >gal> glc>0 --- CHECK ABILITY TO FORM PRODUCT --- be formed --- =0.2 some --- CALCULATE LAG PERIOD MAXIMUM GROWTH /2.5) t+(1.2- 0.016* --- _CYCLE --- Time range 0-45 hours, plot 2950-6550 i.e. times 80 --- Biomass A4160-960 i.e. --- Product i3360-1110 i.e. --- 4060-1560 i.e. --- I>45 x=4.5 >=2.2 --- HIGH ETHANOL TOXIC --- DECELERATION PHASE s<=0.2 --- ACCELERATION --- START EXPONENTIAL --- x+2*u* s-2.5*u/ --- concentration glc/gal s>1.4/ +10*u* --- p+320*u* --- DRAW DATA POINTS 2900+t*80 ,4510-x*800 +100,4610-x*800 60,50,100 ,4110-p*5 +100,4210-p*5 0,50,100 --- ,4010-s*500 +100,4110-s*500 --- 300,50,100 /"fermenter" ,90-x*10,100 t>45 --- END OF --- ADJUST GLC, ETOH FIGURES --- IF NO LEFT, WAIT UNTIL TADD+2 OR T=45 --- OR IF BIOMASS IS AT --- IF STRAIN IS PROTEASE +VE, THEN DECREASE CONCN --- IF $CEASE glc=0 9 =0 gal=0 f =0 t<45) x=4 OR } >=2.2 --- ~ degraded 2900+t*80 ,4510-x*800 +100,4610-x*800 60,50,100 ,4110-p*5 +100,4210-p*5 0,50,100 --- ,4010-s*500 +100,4110-s*500 --- 300,50,100 --- IF T= , ADD SECOND LOT OF glc+2 gal+2 Zt>45 (p/2) buttonup buttonup arrow yield 0 units ethanol 0 mg/ml 333333 ffffff medium fermenter yield ethanol substrate factor substrate1 substrate2 promoter protease culturecolor final MCBcoursewaree courseware MCBBBBBBeware bowtie congratulations Congratulations! You have managed to produce a maximum yield of 274 units of HGH ..est experiment. You have managed to produce a maximum yield of 194 units of HGH. Not bad, but you should really be able to produce over 200 units. You have only managed to produce a maximum yield of 0 units of HGH. You should really aim to produce over 200 units......er 200 units. You haven't performed any experiments! I suggest you go back and try again. Ask your tutor or demonstrator for help if you're having trouble. You haven't performed enough experiments! I suggest you go back and do some more. Ask your tutor or demonstrator for help if you're having trouble. mouth mouth (A) MATa, ura3s Ethanol Ethanol 2 hourshours 0 fly units/ml flyconstant 44 mg/ml exptno Albert's no 3 newgraph arrow arrow SUBSTRATE 2 checkprint "fly" "bum" buttonUp buttonUp bkgd1 letterpage Scottish Biotech PERSONNEL SECTION, Aberdeen Science Park, Aberdeen, AB1 4UR PRIVATE AND CONFIDENTIAL A. Graduate School of Biotechnology Rockall City University Outer Hebrides Dear Graduate Re: Scientific Officer in the Yeast Biotechnology Section On the recommendation of Dr Rust, I am writing to offer you an appointment as Scientific Officer in the Yeast Biotechnology Section, commencing 1 Nov 1993. Your salary will commence at the starting point of the Scientific Officer scale, currently 14200 per annum. The other conditions of your employment are shown on the attached pages. Please confirm, by clicking on the Accept Offer button that you have read and understood the conditions attached to this appointment and that you accept it on the terms stated. Yours sincerely I. Hiremmmemmmmm accept "aims" buttonUp buttonUp Accept Offer decline "We are sorry you have decided decline our offer employment fthe company. Dr Rust has asked us enquire whether bwish reconsider." f"Accept Offer" "Really "aims" "Your mark tutorial 0%. Do want f"Yes" "&No" "letterpage" buttonUp buttonUp We are sorry you have decided to decline our offer of employment with the company. Dr Rust has asked us to enquire whether you wish to reconsider. Accept Offer Really decline offer! Accept Offer Really decline offer! Your mark for this tutorial is 0%. Do you want to try again? letterpage Decline Offer ExitProgram buttonUp buttonUp fedbatch sliderPosition "addglu" "graph1" "etoh1" "graph2" "graph3" "graph4" "graph5" "graph6" "etoh2" "biomass" leavePage leavePage addglu sliderPosition graph1 etoh1 graph2 graph3 graph4 graph5 graph6 etoh2 biomass 0.5237631476431632 Fed-batch culture Fed batch culture, like normal batch culture, involves normal growth of the cells until the substrate or some other medium component is exhausted. The difference is that not all of the growth substrate is added at the start of the experiment... graph1 glucgraph1 glucgraph2 glucgraph3 addglu sliderPosition width glide line id 118 of page id 15 txClassName hSliderThumb 2nd addition of glucose b : _ biomass hintbutton "The concentrations glucose used many the experiments " &\ practical/tutorial are xoptimal maximising cell biomass " &\ Hfed-batch strategies. When )a high , " &\ ethanol production increased." buttonUp buttonUp The concentrations of glucose used in many of the experiments in this practical/tutorial are not optimal for maximising cell biomass by fed-batch strategies. When glucose is at a high concentration, the rate of ethanol production is increased. toxic Ethanol is toxic at a concentration of 30-40mg/ml (depending on conditions present) which will cause the cells to stop growing. At high glucose concentrations, the concentration of ethanol will build up quicker.. biomass Here the limiting factor is something other than the carbon source, such as nitrogen, phosphate, etc. biomass Maximum biomass "biomass" mouseEnter mouseLeave mouseEnter biomass mouseLeave biomass graph2 glucgraph1 glucgraph2 glucgraph3 graph6 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph1 etoh1 Ethanol toxicity "toxic" mouseEnter mouseLeave mouseEnter toxic mouseLeave toxic etoh2 Ethanol toxicity "toxic" mouseEnter mouseLeave mouseEnter toxic mouseLeave toxic graph3 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 graph4 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 graph5 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 Drag the red slider (below the graph) to change the time at which the second batch is added to the culture and see the effect on the growth curve. red slider "helppage" buttonUp buttonUp helppage 82429649 0.5603980782429649 "vacuole" "yeasttxt" "Vacuoles - site storage some metabolites a major macromolecular degradation." mouseEnter mouseLeave mouseEnter vacuole yeasttxt Vacuoles - site of storage of some metabolites and a major site of macromolecular degradation. yeasttxt mouseLeave vacuole yeasttxt yeasttxt "mitochondrion" "yeasttxt" "Mitochondrion - site energy generation Haerobic respiration." mouseEnter mouseLeave mouseEnter mitochondrion yeasttxt Mitochondrion - site of energy generation by aerobic respiration. yeasttxt mouseLeave mitochondrion yeasttxt yeasttxt "yeasttxt" "Lipid - storage vesicle." mouseEnter mouseLeave mouseEnter yeasttxt Lipid - storage vesicle. yeasttxt mouseLeave yeasttxt yeasttxt U"er" "yeasttxt" "Rough endoplasmic reticulum - site translation mRNAs including secreted proteins vacuolar U"er" mouseEnter mouseLeave mouseEnter yeasttxt Rough endoplasmic reticulum - site of translation of mRNAs including secreted proteins and vacuolar proteins. yeasttxt mouseLeave yeasttxt yeasttxt m"cellwall" "yeasttxt" "Thick - comprised mainly proteins carrying large glucan mannan side-chains." mouseEnter mouseLeave mouseEnter cellwall yeasttxt Thick cell wall - comprised mainly of proteins carrying large glucan and mannan side-chains. yeasttxt mouseLeave cellwall yeasttxt yeasttxt "nucleus" "yeasttxt" "Nucleus - site chromosome replication segregation, gene transcription, RNA processing." mouseEnter mouseLeave mouseEnter nucleus yeasttxt Nucleus - site of chromosome replication and segregation, gene transcription, and RNA processing. yeasttxt mouseLeave nucleus yeasttxt yeasttxt nucleolus "nucleolus" "yeasttxt" "Nucleolus - site rRNA transcription ribosome biogenesis." mouseEnter mouseLeave mouseEnter nucleolus yeasttxt Nucleolus - site of rRNA transcription and ribosome biogenesis. yeasttxt mouseLeave nucleolus yeasttxt yeasttxt yeasttxt mouseEnter mouseLeave mouseEnter yeasttxt mouseLeave yeasttxt saccharomyces diauxicgrowth Ulacgraph1 mlacgraph2 Ulacgraph3 mlacgraph4 mlacgraph5 lacexpl labelenz labellac Uglucgraph1 mglucgraph2 Uglucgraph3 labelgluc labalexhaust leavePage leavePage labalexhaust labelgluc glucgraph3 glucgraph2 glucgraph1 labellac labelenz lacexpl lacgraph5 lacgraph4 lacgraph3 lacgraph2 lacgraph1 Diauxic growth glucgraph1 glucgraph2 glucgraph3 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 Growth substrate: 4looptime Uglucgraph1 mglucgraph2 Uglucgraph3 Ulacgraph1 H/15 mlacgraph2 c/15 Ulacgraph3 ~/15 mlacgraph4 mlacgraph5 labelenz labellac lacexpl buttonUp buttonUp lacexpl labellac labelenz lacgraph5 lacgraph4 lacgraph3 lacgraph2 lacgraph1 glucgraph3 glucgraph2 glucgraph1 looptime Glucose + lactose 4looptime Ulacgraph1 mlacgraph2 Ulacgraph3 mlacgraph4 mlacgraph5 labelenz labellac Uglucgraph1 mglucgraph2 Uglucgraph3 buttonUp buttonUp glucgraph3 glucgraph2 glucgraph1 labellac labelenz lacgraph5 lacgraph4 lacgraph3 lacgraph2 lacgraph1 looptime Glucose only labelgluc Growth on glucose labelenz Enzyme induction labalexhaust Glucose exhausted labellac Growth on lactose The graph shows batch growth by E.coli in the presence of glucose. Diauxic growth occurs when E.coli is grown in the presence of glucose and lactose . Click the blue button above to see this. Glucose is more readily catabolised and is used first: once it is exhausted, the enzymes required for lactose catabolism are induced and growth recommences using lactose. lacexpl Glucose is more readily catabolised and is used first; once it is exhausted, the enzymes required for lactose catabolism are induced and growth recommences using lactose. ln biomass fermenter "componentstxt" "arial" leavePage leavePage componentstxt arial componentstxt components componentstxt 444Z4 4j9J<P= =B>H@p@~B bubble Try moving the cursor over the fermenter! The Fermenter vessel Smaller vessels like this one are usually sterilised in an autoclave....................... components baffles These help to improve the efficiency of mixing. stirrer This maintains the homogeneity of the culture. This senses the pH of the culture. pHcontrol This controls the addition of acid and/or alkali to keep the culture pH at the correct value. stirrercontrol This controls the stirring rate and thus the aeration of the culture. settemp This keeps the temperature of the culture at a constant value. tempprobe This senses the temperature of the culture. autoclave This turns brown if the fermentation vessel has been sterilised correctly. injection This can be used to inject cells or nutrients. antifoam This controls foaming of the culture. "components" "componentstxt" "Fermentation vessel" mouseEnter mouseLeave mouseEnter components componentstxt Fermentation vessel components vessel componentstxt mouseLeave components componentstxt components componentstxt pH electrode "components" "componentstxt" "pH electrode" "ph" mouseEnter mouseLeave mouseEnter components componentstxt pH electrode components componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "Temperature probe" "tempprobe" mouseEnter mouseLeave mouseEnter components componentstxt Temperature probe components tempprobe componentstxt mouseLeave components componentstxt components componentstxt "components" "temperature display" mouseEnter mouseLeave mouseEnter components temperature display components mouseLeave components components "components" "componentstxt" "Temperature control module" "settemp" mouseEnter mouseLeave mouseEnter components componentstxt Temperature control module components settemp componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "pH control module" "phcontrol" mouseEnter mouseLeave mouseEnter components componentstxt pH control module components phcontrol componentstxt mouseLeave components componentstxt components componentstxt speed "components" "pH display" mouseEnter mouseLeave mouseEnter components pH display components mouseLeave components components pHdisplay speeddisplay "components" "speed display ( rpm)" mouseEnter mouseLeave mouseEnter components speed display (in rpm) components mouseLeave components components speed "components" "on/off switch" mouseEnter mouseLeave mouseEnter components on/off switch components mouseLeave components components speedknob "components" "componentstxt" "Stirrer control module" "stirrercontrol" "bar" "centre" ouseEnter mouseLeave mouseEnter buttonUp mouseEnter components componentstxt Stirrer control module components stirrercontrol componentstxt mouseLeave components componentstxt components componentstxt buttonUp centre centre right right "components" "componentstxt" "Antifoam inlet" "antifoam" mouseEnter mouseLeave mouseEnter components componentstxt Antifoam inlet components antifoam componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "Injection port" "injection" mouseEnter mouseLeave mouseEnter components componentstxt Injection port components injection componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "Outlet port" "outlet" mouseEnter mouseLeave mouseEnter components componentstxt Outlet port components outlet componentstxt mouseLeave components componentstxt components componentstxt rbaffle "components" "componentstxt" "Baffles" "baffles" mouseEnter mouseLeave mouseEnter components componentstxt Baffles components baffles componentstxt mouseLeave components componentstxt components componentstxt rbaffle "components" "componentstxt" "Baffles" "baffles" mouseEnter mouseLeave mouseEnter components componentstxt Baffles components baffles componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "Stirrer" "stirrer" mouseEnter mouseLeave mouseEnter components componentstxt Stirrer components stirrer componentstxt mouseLeave components componentstxt components componentstxt right centre "bar" "centre" "components" "componentstxt" "Cold finger" "coldfinger" mouseEnter mouseLeave mouseEnter components componentstxt Cold finger components coldfinger componentstxt mouseLeave components componentstxt components componentstxt coldfinger This maintains the temperature of the culture at the correct value. Water is continuously circulated through it from a constant-temperature water bath. componentstxt outlet This allows removal of samples for testing. "components" "componentstxt" "Acid/base inlet" "acid" mouseEnter mouseLeave mouseEnter components componentstxt Acid/base inlet components componentstxt mouseLeave components componentstxt components componentstxt "components" "componentstxt" "Autoclave tape" "autoclave" mouseEnter mouseLeave mouseEnter components componentstxt Autoclave tape components autoclave componentstxt mouseLeave components componentstxt components componentstxt This allows addition of acid and/or base for control of pH. fedbatch sliderPosition "addglu" "graph1" "etoh1" "graph2" "graph3" "graph4" "graph5" "graph6" "etoh2" "biomass" leavePage leavePage addglu sliderPosition graph1 etoh1 graph2 graph3 graph4 graph5 graph6 etoh2 biomass 0.5237631476431632 Fed-batch culture Fed batch culture, like normal batch culture, involves normal growth of the cells until the substrate or some other medium component is exhausted. The difference is that not all of the growth substrate is added at the start of the experiment... graph1 glucgraph1 glucgraph2 glucgraph3 addglu sliderPosition width glide line id 118 of page id 15 txClassName hSliderThumb 2nd addition of glucose b : _ biomass hintbutton "The concentrations glucose used many the experiments " &\ practical/tutorial are xoptimal maximising cell biomass " &\ Hfed-batch strategies. When )a high , " &\ ethanol production increased." buttonUp buttonUp The concentrations of glucose used in many of the experiments in this practical/tutorial are not optimal for maximising cell biomass by fed-batch strategies. When glucose is at a high concentration, the rate of ethanol production is increased. toxic Ethanol is toxic at a concentration of 30-40mg/ml (depending on conditions present) which will cause the cells to stop growing. At high glucose concentrations, the concentration of ethanol will build up quicker.. biomass Here the limiting factor is something other than the carbon source, such as nitrogen, phosphate, etc. biomass Maximum biomass "biomass" mouseEnter mouseLeave mouseEnter biomass mouseLeave biomass graph2 glucgraph1 glucgraph2 glucgraph3 graph6 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph1 etoh1 Ethanol toxicity "toxic" mouseEnter mouseLeave mouseEnter toxic mouseLeave toxic etoh2 Ethanol toxicity "toxic" mouseEnter mouseLeave mouseEnter toxic mouseLeave toxic graph3 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 graph4 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 graph5 lacgraph1 lacgraph2 lacgraph3 lacgraph4 lacgraph5 Drag the red slider (below the graph) to change the time at which the second batch is added to the culture and see the effect on the growth curve. red slider "helppage" buttonUp buttonUp helppage foreigngene "foreigntxt" "joketxt" "promoterbuts" "adhtxt" "galtxt" "pgktxt" "genotype" enterPage enterPage foreigntxt joketxt promoterbuts adhtxt galtxt pgktxt genotype z!<,b- foreigntxt An expression system is dependent on: The genetic make-up of the host yeast cell The stability and copy-number of the vector The transcriptional promoter The presence or absence of a signal sequence The nature of the foreign sequence sequence ature of the foreign sequence Foreign gene expression systems t"$ ~ promoterbuts adhtxt galtxt joketxt pgktxt buttonUp buttonUp pgktxt joketxt galtxt adhtxt pgktxt galtxt joketxt adhtxt buttonUp buttonUp adhtxt joketxt galtxt pgktxt adhtxt pgktxt joketxt galtxt buttonUp buttonUp galtxt joketxt pgktxt adhtxt PROMOTER: try clicking on the buttons below pgktxt REGULATION OF THE PGK1 GENE CARBON SOURCE ACTIVITY GLUCOSE ETHANOL NEITHERRR NEITHER CELLS AREN'T GROWING! ETHANOL ETHANOL NEITHERRR ETHANOL NEITHERRR galtxt REGULATION OF THE GAL1 GENE CARBON SOURCE ACTIVITY GLUCOSE ETHANOL NEITHERRR BINGO! GALACTOSE HANOL NEITHERRR ETHANOL NEITHERRR NEITHER ETHANOL NEITHERRR ~ V { j!B!g! adhtxt b#:#_# REGULATION OF THE ADH2 GENE CARBON SOURCE \$4$Y$ ACTIVITY GLUCOSE ETHANOL NEITHERRR BINGO! ETHANOL ETHANOL NEITHERRR ETHANOL NEITHERRR p*H*m* NEITHER WHAT'S A CELL TO GROW ON?? promoter "joketxt" "foreigntxt" "genotype" "promoterbuts" buttonUp buttonUp joketxt foreigntxt genotype promoterbuts Promoter strain "galtxt" "adhtxt" "pgktxt" "promoterbuts" "joketxt" "foreigntxt" "genotype" buttonUp buttonUp galtxt adhtxt pgktxt promoterbuts joketxt foreigntxt genotype Strain genotype GENOTYPES OF CELLS USED IN THIS TUTORIAL MATa Mating type a. These haploid cells will mate with haploid cells of the opposite mating type [MATa] to form diploids. Mating type will not affect HGH production in your experiments. ura3 This mutation imposes an auxotrophic requirement for uracil. The selectable marker used on the plasmid in your experiments is the URA3 gene. pep4 This mutation disrupts the PEP4 gene which is important for the activation of vacuolar proteases. prb1 The prb1 also is important for the activation of vacuolar proteases. Therefore prb1 mutants have reduced vacuolar protease activity. joketxt Click on one of the buttons on the right for further inform-ation.. 4:PHYSSIZE We like to think of ourselves as yeast molecular biologists....=6 fedbatch letterpage startpage bkgd2 0 10 20 30 40 Time (hours)))rs)e (hours)hours)rs)urs)urs) Biomass Productucttttroductt - 0.66 - 1.66 - 2.66 0.4446 l D i ln biomass (ln mg/ml)])) Final HGH concentration: Final ethanol concentration: speed speedknob rbaffle rbaffle "bubble" "components" "stirrer" mouseEnter mouseLeave mouseEnter bubble components stirrer components mouseLeave components components right centre "bar" "centre" buttonup buttonup centre centre right right centre centre right right Dialog ctrlID dlgBox 524480,22,26,24,307,192,,,Plan experiment,8,Helv,,7.62,9.68,18.29,19.69,63,1342177283,130,toolbook,0,promoter,33.41,11.16,62.10,54.28,16,1342177287,128, Promoter,0,strain,106.40,10.95,140.15,54.28,29,1342177287,128, Yeast strain,0,substrate1,33.60,75.49,61.87,54.28,33,1342177287,128, Substrate 1,0,substrate2,106.40,75.49,140.15,54.28,37,1342177287,128, Substrate 2 ,0,addedafter,175.20,88.74,44.80,11.45,51,1342177280,130,Added after:,0,,7.62,136.33,293.07,36.18,182,1342177280,130,_________________________________________________________________________ Plan your experiment by clicking on your choice of promoter,0,ok,255.73,14.56,45.33,16.33,12,1342242817,128,&OK,0,cancel,255.85,40.90,44.80,16.08,14,1342242816,128,&Cancel,0,adh,40.80,23.84,42.90,10.05,21,1342308361,128,ADH2,0,gal,40.80,35.90,41.07,10.05,22,1342177289,128,GAL1,0,pgk,40.80,48.98,41.07,10.05,171,1342177289,128,PGK1,0,straina,114.25,23.84,125.10,10.46,40,1342308361,128,Strain A: MATa ura3,0,strainb,114.25,36.64,123.62,10.46,39,1342177289,128,Strain B: MATa ura3 pep4 prb1,0,strainc,114.25,49.64,125.10,10.46,38,1342177289,128,Strain C: MATa ura3 pep4,0,ethanol,41.22,89.15,50.90,10.75,41,1342308361,128,Ethanol,0,galactose,40.80,101.78,50.90,10.75,43,1342177289,128,Galactose,0,glucose,41.03,114.58,50.93,10.75,42,1342177289,128,Glucose,0,ethanol2,114.25,89.15,50.93,11.16,46,1342308361,128,Ethanol,0,galactose2,114.25,102.73,50.93,11.12,44,1342177289,128,Galactose,0,glucose2,114.25,115.57,50.93,11.16,45,1342177289,128,Glucose,0,tadd,174.63,100.80,59.43,54.31,227,1352859715,133,,0 <$dlgInit ,h63, groupbox promoter,g16, Promoter groupbox strain,g29, Yeast strain groupbox substrate1,g33, Substrate 1 groupbox substrate2,g37, Substrate 2 static addedafter,s51,Added after: ,s182,_________________________________________________________________________ Plan your experiment by clicking on your choice of promoter, yeast strain, substrates and the time you want to add the second substrate, then click OK to save these settings. button ok,b12,TRUE button cancel,b14,FALSE button adh,b21,TRUE button gal,b22,FALSE button pgk,b171,FALSE button straina,b40,TRUE button strainb,b39,FALSE button strainc,b38,FALSE button ethanol,b41,TRUE button galactose,b43,FALSE button glucose,b42,FALSE button ethanol2,b46,TRUE button galactose2,b44,FALSE button glucose2,b45,FALSE combobox tadd,c227,2 hours 4 hours 6 hours &8 hours 10 hours 12 hours 14 hours 16 hours 18 hours 20 hours 22 hours 24 hours 26 hours 28 hours 30 hours 32 hours 34 hours 36 hours 38 hours 40 hours Plan experiment clear 4culturecolor "medium" ,90,100 buttonup buttonup medium culturecolor Clear graph inoculate Inoculate concn (Units/ml)4 INITIALISE 4protease,substrate1,substrate2,promoter,tadd,planinit GET SETTINGS FROM DIALOG BOX dlginit retvalue dialog(dlgbox MODIFY getvalue( BOK") Bstraina") "(A)" "MATa, ura3" Bstrainb") "(B)" e, pep4, prb1" Bstrainc") "(C)" Badh") "ADH2" Bgal") "GAL1" Bpgk") "PGK1" Bethanol") "Ethanol" Bgalactose") "Galactose" Bglucose") "Glucose" Bethanol2") Bgalactose2") Bglucose2") ,"combobox --- "&"& buttonup buttonup dlginit dlgbox dialog button OK getvalue button straina getvalue protease protease MATa, ura3 button strainb getvalue ffffff protease protease MATa, ura3, pep4, prb1 button strainc getvalue 333333 protease protease MATa, ura3, pep4 button adh getvalue promoter button gal getvalue promoter button pgk getvalue promoter button ethanol getvalue substrate1 Ethanol button galactose getvalue substrate1 Galactose button glucose getvalue substrate1 Glucose button ethanol2 getvalue substrate2 Ethanol button galactose2 getvalue substrate2 Galactose button glucose2 getvalue substrate2 Glucose combobox tadd getvalue retvalue protease substrate1 substrate2 promoter planinit saccharomyces enterPage enterPage ,8.2/ Saccharomyces cerevisiae beericon beertxt S.cerevisiae is very important commercially because of its use in the brewing, wine-making and bread-making industries.... Point to different parts of the yeast cell or to one of the icons on the right to learn more about them. beertxt mouseEnter mouseLeave mouseEnter beertxt mouseLeave beertxt biotechtxt mouseEnter mouseLeave mouseEnter biotechtxt mouseLeave biotechtxt :PHYSSIZE yeasttxt yeasttxt Yeasts are important scientific tools. By studying the genes controlling cell division in these simple eucaryotic organisms, insights are being obtained into human diseases which affect cell division e.g. cancer.............cer. biotechtxt Yeasts are important tools in biotechnology. For example, they can be used in foreign gene expression systems in the production of valuable proteins. lipid "lipid" "yeasttxt" "Lipid" mouseEnter mouseLeave mouseEnter lipid yeasttxt Lipid yeasttxt mouseLeave lipid yeasttxt yeasttxt nucleus vacuole cellwall mitochondrion lipid "vacuole" "yeasttxt" "Vacuoles - site storage some metabolites a major macromolecular degradation." mouseEnter mouseLeave mouseEnter vacuole yeasttxt Vacuoles - site of storage of some metabolites and a major site of macromolecular degradation. yeasttxt mouseLeave vacuole yeasttxt yeasttxt "mitochondrion" "yeasttxt" "Mitochondrion - site energy generation Haerobic respiration." mouseEnter mouseLeave mouseEnter mitochondrion yeasttxt Mitochondrion - site of energy generation by aerobic respiration. yeasttxt mouseLeave mitochondrion yeasttxt yeasttxt "yeasttxt" "Lipid - storage vesicle." mouseEnter mouseLeave mouseEnter yeasttxt Lipid - storage vesicle. yeasttxt mouseLeave yeasttxt yeasttxt U"er" "yeasttxt" "Rough endoplasmic reticulum - site translation mRNAs including secreted proteins vacuolar U"er" mouseEnter mouseLeave mouseEnter yeasttxt Rough endoplasmic reticulum - site of translation of mRNAs including secreted proteins and vacuolar proteins. yeasttxt mouseLeave yeasttxt yeasttxt m"cellwall" "yeasttxt" "Thick - comprised mainly proteins carrying large glucan mannan side-chains." mouseEnter mouseLeave mouseEnter cellwall yeasttxt Thick cell wall - comprised mainly of proteins carrying large glucan and mannan side-chains. yeasttxt mouseLeave cellwall yeasttxt yeasttxt "nucleus" "yeasttxt" "Nucleus - site chromosome replication segregation, gene transcription, RNA processing." mouseEnter mouseLeave mouseEnter nucleus yeasttxt Nucleus - site of chromosome replication and segregation, gene transcription, and RNA processing. yeasttxt mouseLeave nucleus yeasttxt yeasttxt nucleolus "nucleolus" "yeasttxt" "Nucleolus - site rRNA transcription ribosome biogenesis." mouseEnter mouseLeave mouseEnter nucleolus yeasttxt Nucleolus - site of rRNA transcription and ribosome biogenesis. yeasttxt mouseLeave nucleolus yeasttxt yeasttxt yeasttxt mouseEnter mouseLeave mouseEnter yeasttxt mouseLeave yeasttxt Nucleus - site of chromosome replication and segregation, gene transcription, and RNA processing. background background button buttonUp buttonUp buttonUp background button Library pH electrode v=N=s= |>T>y> speed &9~@, speedknob ~ > > + questions2 "studentname" "exp1" "exp2" "exp3" B"printgraphbut" 4nosave,checkgraph ZmyList "0123456789" M = 13) "0123456789" "You must type enterPage keyChar enterPage studentname printgraphbut keyChar 0123456789 buttonUp printgraphbut 0123456789 You must type in a number. myList nosave checkgraph printgraphbut PRINT GRAPH Type the experiment numbers you wish to print in the red boxes below. When you are sure these are the graphs you want to keep, click ONCE on the "PRINT GRAPH" button below or press the "Enter" key. Optimal conditions for GAL1 promoter - experiment number Optimal conditions for ADH2 promoter - experiment number Optimal conditions for PGK1 promoter - experiment number Type your name and your partner's name in the red box below. Button printing printing Printing.... Be patient! } H | run button 4inoc "medium" "autoclave tape" buttonUp buttonUp medium autoclave tape buttonUp buttonUp buttonUp run button buttonUp buttonUp buttonUp run button Laboratory Strain: Promoter: Substrate 1: Substrate 2: after medium PRFRcolor dRfillcolour 0,50,100 autoclave tape protease hS@SeS (A)MATa,ura3a3, pep4, prb1 promoter ADH2ose substrate1 rTJToT Ethanolse substrate2 Ethanolse xUPUuU 8 hourss hours Dialog UctrlID UdlgBox 524480,22,26,24,307,192,,,Plan experiment,8,Helv,,7.62,9.68,18.29,19.69,63,1342177283,130,toolbook,0,promoter,33.41,11.16,62.10,54.28,16,1342177287,128, Promoter,0,strain,106.40,10.95,140.15,54.28,29,1342177287,128, Yeast strain,0,substrate1,33.60,75.49,61.87,54.28,33,1342177287,128, Substrate 1,0,substrate2,106.40,75.49,140.15,54.28,37,1342177287,128, Substrate 2 ,0,addedafter,175.20,88.74,44.80,11.45,51,1342177280,130,Added after:,0,,7.62,136.33,293.07,36.18,182,1342177280,130,_________________________________________________________________________ Plan your experiment by clicking on your choice of promoter,0,ok,255.73,14.56,45.33,16.33,12,1342242817,128,&OK,0,cancel,255.85,40.90,44.80,16.08,14,1342242816,128,&Cancel,0,adh,40.80,23.84,42.90,10.05,21,1342308361,128,ADH2,0,gal,40.80,35.90,41.07,10.05,22,1342177289,128,GAL1,0,pgk,40.80,48.98,41.07,10.05,171,1342177289,128,PGK1,0,straina,114.25,23.84,125.10,10.46,40,1342308361,128,Strain A: MATa ura3,0,strainb,114.25,36.64,123.62,10.46,39,1342177289,128,Strain B: MATa ura3 pep4 prb1,0,strainc,114.25,49.64,125.10,10.46,38,1342177289,128,Strain C: MATa ura3 pep4,0,ethanol,41.22,89.15,50.90,10.75,41,1342308361,128,Ethanol,0,galactose,40.80,101.78,50.90,10.75,43,1342177289,128,Ga 4checkgraph, nosave ZmyList "exp1" "exp2" "exp3" "Remember type a EACH box." "You have xperformed many experiments!" "Experiment 0!?! ygotta be kiddin'!." "studentname" your xsaved one these 8." & \ B"printgraphbut" "Are sure want p graphs " && ( &&","&& ( ) &&" "&& ( ) &&"?" f"Print" "printing" you don't printed /"labbk "hideprint" --match the chosen correct labbook /" + 1 "checkprint" "studname" "(" & & ") Name:" && 1000,1000,1000,1000 " = "" "bar" 3195,4560,3250,4755 3195,4560,3250+a,4755 Za=2200 3195,4560,3250,4755 "questions2" eatclicks() buttonUp buttonUp continue Remember to type a number into EACH box. You have not performed this many experiments! Experiment 0!?! You gotta be kiddin'!. studentname Remember to type in your name! studentname You have not saved one of these experiments in your lab book. Go back and check. printgraphbut Are you sure you want to print the graphs from experiments Print printgraphbut printing labbk background 0,100,0 hideprint labbk background labbk background checkprint check studname studname ) Name: studentname checkprint checkprint text of field "checkprint" = "check" labbk background labbk background checkprint studname studname studname printing printing buttonup continue studentname questions2 questions2 questions2 questions2 eatclicks myList checkgraph nosave Biomass Product 0 10 20 30 40 Time (hours)))rs)e (hours)hours)rs)urs)urs) - 0.66 - 1.66 - 2.66 0.4446 ln biomass (ln mg/ml)])) concn (Units/ml)4 Final HGH concn: Final ethanol concn:tration::::::::: yield 0 units ethanol 38 mg/ml labtxt RUNNING THE EXPERIMENT Use the yellow buttons on the right to plan, prepare and start an experiment. Your chosen parameters will appear on the grey bar above. Remember to keep an eye on the fermenter!fermentor display on the left. arrow arrow x P u SUBSTRATE 2 ExitProgram buttonUp buttonUp help button "helppage" buttonUp buttonUp helppage q button "questions1" buttonUp buttonUp questions1 buttonUp buttonUp buttonUp q button Report aims button "aims" buttonUp buttonUp target B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button results button "results1" buttonUp buttonUp results1 B"results buttonUp buttonUp buttonUp results button LabBook saccharomyces "saccharomyces" buttonUp buttonUp saccharomyces Saccharomyces foreign "foreigngene" buttonUp buttonUp foreigngene Foreign gene expression systems fedbatch "fedbatch" buttonUp buttonUp fedbatch Fed-batch culture fermenter "fermenter" buttonUp buttonUp fermenter The fermenter diauxic "diauxicgrowth" buttonUp buttonUp diauxicgrowth Diauxic growth LIBRARY run button "medium" /"fermenter" "autoclave tape" inoc buttonUp buttonUp medium fermenter autoclave tape fermenter buttonUp buttonUp buttonUp run button Laboratory buttonUp buttonUp buttonUp run button buttonUp buttonUp background background button buttonUp buttonUp buttonUp background button Library buttonUp buttonUp buttonUp q button B"results buttonUp buttonUp buttonUp results button f727C7 helppage "info" "scrollbarinfo" "hotwordfound" "menubarinfo" enterpage leavepage enterpage - 2 - - 1 - leavepage scrollbarinfo hotwordfound menubarinfo "info" buttonup buttonup - 2 - - 1 - How to use this program "info" buttonup buttonup - 5 - - 4 - Buttons and scroll bars "info" buttonup buttonup - 6 - - 5 - Push button "info" buttonup buttonup - 8 - - 7 - Checkbox "info" buttonup buttonup - 7 - - 6 - Rounded button "info" buttonup buttonup - 9 - - 8 - Radio button "info" buttonup buttonup - 11 - - 10 - Buttons at the foot of the pagethe page ExitProgram "info" buttonup buttonup - 12 - - 11 - Other featuress at the foot of the page Hotword Hotword "info" buttonup buttonup - 14 - - 13 - "info" buttonup buttonup - 15 - - 14 - Menubarem "info" buttonup buttonup - END - - 15 - Windows "info" buttonup buttonup - 10 - - 9 - Scroll bars - - - - - - - 1 - - - - - - - HOW TO USE THIS PROGRAM This page contains a number of terms and items that you may not be familiar with if you have not used a similar program before. Just click on one of the objects in the blue boxes on the right for further information... - - - - - - - 2 - - - - - - - THE MOUSE Use this to move the cursor to where you want on the page. Events may happen by simply moving the cursor over an object, or you may need to click or double click the left hand side of the mouse. Be patient, sometimes this takes time to work! - - - - - - - 3 - - - - - - - DRAGGING To drag an object, position the cursor over the object, then, while holding the left mouse button down, move the cursor to the position where you want the object to be and release the mouse button. - - - - - - - 4 - - - - - - - BUTTONS AND SCROLL BARS Buttons, as their name suggests, are items that you can "push" by clicking on them using the left button on the mouse. Scroll bars help you move around text. - - - - - - - 5 - - - - - - - PUSH BUTTONS Push buttons, as their name suggests, can be "pushed" by clicking on the button using the left button on the mouse. The word on the push button will indicate what will happen when you press it. - - - - - - - 6 - - - - - - - ROUNDED BUTTONS Rounded buttons are identical in use to push buttons. To "push" the button, simply click on it. The word on the button will indicate what will happen when you press it. - - - - - - - 7 - - - - - - - CHECKBOXES Checkboxes are a special type of button. A cross in the box indicates that you have selected the option on the button label. Test this by clicking on the button a few times... - - - - - - - 8 - - - - - - - RADIO BUTTONS Radio buttons are a special type of button. A dot in the circle indicates that you have selected the option on the button label. Test this by clicking on the button a few times... - - - - - - - 9 - - - - - - - SCROLL BARS Scroll bars are found at the right hand side of some boxes containing text. Click on to move upwards and to move downwards through the text. To move more quickly, keep the mouse button pressed down. - - - - - - 10 - - - - - - BUTTONS WITH ICONS These buttons are at the foot of every page. The icon and words vary according to the sections in the tutorial. Click on one to go to a new section. - - - - - - 11 - - - - - - THE EXIT BUTTON This button is on the far left at the foot of each page. Click on it when you wish to end the program. As a precaution, you will be asked whether you really want to quit before the tutorial ends. - - - - - - 12 - - - - - - THE HELP BUTTON This button is at the foot of every page. Click on it to move back to the this page at any time. - - - - - - 13 - - - - - - HOTWORDS Hotwords are found buried in normal text and are either in a bold typeface or are highlighted by a box around the word. When you click on the hotword, you will get more information on that word. - - - - - - 14 - - - - - - THE MENUBAR The menubar is the bar along the top of the page just below the title. If you click on any of the words there, a drop-down menu will appear. Click a word or phrase on the drop-down part to choose an option e.g. to print your data or to get more help. - - - - - - 15 - - - - - - WINDOWS Windows is the system that this computer uses to run programs. Each program runs in a separate "window" with a border. - - - - - END - - - - ----------he drop-down part to choose an option e.g. to print your data or to get more help. - - - - - - 16 - - - - - - WINDOWS Windows is the system that this computer uses to run programs. Each program runs in a separate "window" with a border. - - - - - END - - - - -more help. - - - - - - 16 - - - - - - WINDOWS Windows is the system that this computer uses to run programs. Each program runs in a separate "window" with a border. - - - - - END - - - - ----- "scrollbarinfo" buttonup buttonup scrollbarinfo "hotwordfound" buttonup buttonup hotwordfound "menubarinfo" buttonup buttonup menubarinfo menubarinfo The menu bar is this bar along the top of the page. Click on OK to hide this box.. "menubarinfo" buttonup buttonup menubarinfo scrollbarinfo Click on this arrow to move upwards through the text. Keep the mouse button pressed down to move more quickly. Hold the mouse button down and drag the small box up and down the scroll bar to move up and down through the text. Click on this arrow to move downwards through the text. Keep the mouse pressed down to move more quickly. Click on OK to hide this box. scrollbarinfo buttonup buttonup scrollbarinfo hotwordfound Yes! You've found a hotword. Click on OK to hide this box. s box. "hotwordfound" buttonup buttonup hotwordfound "info" buttonup buttonup - 11 - - 10 - aims button target help button "info" 13 -" buttonup buttonup - 13 - - 12 - "info" buttonup buttonup - 3 - - 2 - Using the mouse "info" buttonup buttonup - 3 - - 2 - "info" buttonup buttonup - 4 - - 3 - N6&6K6 Dragging buttonUp buttonUp Click here to return to page you left background final --- DECLARE SYSTEM VARIABLES --- Enter fthese --- Substrate: 10=Glc 7=Gal 5=EtOH --- substrate1= ; substrate2= " values also serve %conversion factors calculating --- lags, growth rates v utilisation --- Promoter: 10=PGK 7=GAL 5=ADH --- tadd= addition , must be a multiple --- protease: 0.65=high 0.85=medium 0.95=low ,promoter, E,culturecolor AND SET LOCAL Zable,t,u,p,x,glc,gal,etoh,s, ,lag,umax, --- Set I(t), (u), product concn (p), biomass (x), --- glucose f), galactose s), ethanol Ps), identity ( --- ( calculated main loop) --- Move arrow 2500+ *80,1000 --- Clear out yield "0 units" "0 mg/ml" --- MAIN LOOP --- Repeat --- SUBSTRATE CONCENTRATION --- Preference >gal> glc>0 --- CHECK ABILITY TO FORM PRODUCT --- be formed --- =0.2 some --- CALCULATE LAG PERIOD MAXIMUM GROWTH /2.5) t+(1.2- 0.016* --- _CYCLE --- Time range 0-45 hours, plot 2950-6550 i.e. times 80 --- Biomass A4160-960 i.e. --- Product i3360-1110 i.e. --- 4060-1560 i.e. --- I>45 x=4.5 >=2.2 --- HIGH ETHANOL TOXIC --- DECELERATION PHASE s<=0.2 --- ACCELERATION --- START EXPONENTIAL --- x+2*u* s-2.5*u/ --- concentration glc/gal s>1.4/ +10*u* --- p+320*u* --- DRAW DATA POINTS 2900+t*80 ,4510-x*800 +100,4610-x*800 60,50,100 ,4110-p*5 +100,4210-p*5 0,50,100 --- ,4010-s*500 +100,4110-s*500 --- 300,50,100 ,90-x*10,100 t>45 --- END OF --- ADJUST GLC, ETOH FIGURES --- IF NO LEFT, WAIT UNTIL TADD+2 OR T=45 --- OR IF BIOMASS IS AT --- IF STRAIN IS PROTEASE +VE, THEN DECREASE CONCN --- IF $CEASE glc=0 gal=0 L =0 t<45) x=4 OR c >=2.2 --- h degraded 2900+t*80 ,4510-x*800 +100,4610-x*800 60,50,100 ,4110-p*5 +100,4210-p*5 0,50,100 --- ,4010-s*500 +100,4110-s*500 --- 300,50,100 --- IF T= , ADD SECOND LOT OF glc+2 gal+2 Zt>45 (p/2) buttonup buttonup arrow yield 0 units ethanol 0 mg/ml 333333 ffffff medium yield ethanol substrate factor substrate1 substrate2 promoter protease culturecolor questions2 "studentname" "exp1" "exp2" "exp3" B"printgraphbut" 4nosave,checkgraph ZmyList "0123456789" M = 13) "0123456789" "You must type enterPage keyChar enterPage studentname printgraphbut keyChar 0123456789 buttonUp printgraphbut 0123456789 You must type in a number. myList nosave checkgraph printgraphbut PRINT GRAPH Type the experiment numbers you wish to print in the red boxes below. When you are sure these are the graphs you want to keep, click ONCE on the "PRINT GRAPH" button below or press the "Enter" key. Optimal conditions for GAL1 promoter - experiment number Optimal conditions for ADH2 promoter - experiment number Optimal conditions for PGK1 promoter - experiment number Type your name and your partner's name in the red box below. Button printing printing Printing.... Be patient! helppage fermenter questions3 4looptime enterPage enterPage looptime bkgd2 jeanandalb 4checkalbert, checkjean "Please don't ignore Albert, he has some useful information Click on his picture on the Shand side screen see what he e Jean, l really need her help! further" objectives "You must look icons on jeanandalb objectives jeanandalb Please don't ignore Albert, he has some useful information for you. Click on his picture on the left hand side of the screen and see what he has to say before you go on Please don't ignore Jean, you really need her help! Click on her picture on the left hand side of the screen before you go any further checkalbert checkjean objectives You must look at all the objectives before you continue.Click on the icons on the right hand side now #D#p# 32405N8@:2< =*?FC \r]F^ ilk lPp sFwD{6 "albertspeak" "jeanspeak" "rustspeak" buttonUp buttonUp albertspeak jeanspeak rustspeak Dr Alistair Rust Head of Researcht "rustspeak" "jeanspeak" "albertspeak" buttonUp buttonUp rustspeak jeanspeak albertspeak Albert Lab Assistantrchf Researcht "albertspeak" "rustspeak" "jeanspeak" buttonUp buttonUp albertspeak rustspeak jeanspeak Jean Banks Librarianrchf Researcht ExitProgram "jeanspeak" "exithint" ouseEnter mouseLeave mouseEnter buttonUp mouseEnter jeanspeak exithint mouseLeave exithint buttonUp q button Report aims button target B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button results button LabBookk run button Laboratory background button buttonUp buttonUp buttonUp background button Library help button "helppage" "jeanspeak" "aims" "helphint" tonUp mouseEnter buttonUp mouseLeave buttonUp helppage mouseEnter jeanspeak helphint mouseLeave jeanspeak helphint 4checkjean "albertspeak" "rustspeak" "jeanspeak" "jeanhint" tonUp mouseEnter buttonUp mouseLeave buttonUp albertspeak rustspeak jeanspeak checkjean mouseEnter jeanspeak jeanhint mouseLeave jeanhint "albertspeak" "jeanspeak" "rustspeak" "rusthint" tonUp mouseEnter buttonUp mouseLeave buttonUp albertspeak jeanspeak rustspeak mouseEnter jeanspeak rusthint mouseLeave rusthint "jeanspeak" "aimshint" mouseEnter mouseLeave mouseEnter jeanspeak aimshint mouseLeave aimshint 4checkalbert, checkjean, objective4, checkalbres "run1" jeanandalb objectives "You must the library 4you working aboratory." "jeanspeak" "aims" "labhint" tonUp mouseEnter buttonUp mouseLeave buttonUp run button run button jeanandalb objectives You must go to the library before you you start working in the laboratory. checkalbert checkjean objective4 checkalbres mouseEnter jeanspeak labhint mouseLeave labhint labhint "jeanspeak" "titlehint" mouseEnter mouseLeave mouseEnter jeanspeak titlehint mouseLeave titlehint ? Objective 1 4view,analysepage,data "showdata" buttonup buttonup buttonup showdata analysepage "jeanspeak" "ob1hint" 4objective1 B"ob1button" ouseEnter mouseLeave mouseEnter buttonUp mouseEnter jeanspeak ob1hint mouseLeave ob1hint buttonUp buttonUp ob1button objective1 Objective 444444444444444 *W[!& `M8M]M Objective 22 CH20H ^O6O[O 0HHHH 0HHHH JP"PGP 0HHHH OHHHH HHHHH HHHHH HHHHH HHHHH HHHHH OOHHH f!\ > l! "jeanspeak" "ob2hint" 4objective1, objective2, usedbefore 5>=1 B"ob2button" "You must read these objectives correct order. Go click on ouseEnter mouseLeave mouseEnter buttonUp mouseEnter jeanspeak ob2hint mouseLeave ob2hint buttonUp buttonUp ob2button You must read these objectives in the correct order. Go back and click on the first objective. objective1 objective2 usedbefore Objective 392? C=10x27? Objective 3 44x92?pep4 10x27???????? "jeanspeak" "ob3hint" 4objective1, objective2, objective3,usedbefore 9>=1 5>=1) B"ob3button" "You must read these objectives correct order. Go click on ouseEnter mouseLeave mouseEnter buttonUp mouseEnter jeanspeak ob3hint mouseLeave ob3hint buttonUp buttonUp ob3button You must read these objectives in the correct order. Go back and click on the first objective. You must read these objectives in the correct order. Go back and click on the first objective. You must read these objectives in the correct order. Go back and click on the second objective. objective1 objective2 objective3 usedbefore albert g:PHYSSIZE 4checkalbert "rustspeak" "jeanspeak" "albertspeak" "alberthint" tonUp mouseEnter buttonUp mouseLeave buttonUp rustspeak jeanspeak albertspeak checkalbert mouseEnter jeanspeak alberthint mouseLeave alberthint 4checkalbres, checkalbert, checkjean, objective4 B"results jeanandalb objectives "You must the library 4you look )Albert's "results1" "jeanspeak" "aims" "resultshint" tonUp mouseEnter buttonUp mouseLeave buttonUp results button results button jeanandalb objectives You must go to the library before you look at Albert's results. results1 checkalbres checkalbert checkjean objective4 mouseEnter jeanspeak resultshint mouseLeave resultshint resultshint 4checkalbert, checkjean, objective4 jeanandalb objectives "questions1" "jeanspeak" "aims" "qhint" tonUp mouseEnter buttonUp mouseLeave buttonUp q button q button jeanandalb objectives questions1 checkalbert checkjean objective4 mouseEnter jeanspeak qhint mouseLeave qhint qhint 4checkalbert,checkjean,objective4,checkalbres jeanandalb objectives "jeanspeak" "aims" "libhint" tonUp mouseEnter buttonUp mouseLeave buttonUp background button background button jeanandalb objectives background checkalbert checkjean objective4 checkalbres mouseEnter jeanspeak libhint mouseLeave libhint libhint ob1button init dlgInit setValue( ,"","") --DLL function retValue dialog(dlgBox getValue( ., "") buttonUp buttonUp dlgInit dlgBox dialog retValue xdlgInit ,b5,FALSE ,h12, ,s13,Albert has already performed some preliminary experiments. Your first objective is to study the results of these experiments and suggest why he has not managed to produce any HGH so far. ydlgBox 524480,3,150,30,135,100,,,OBJECTIVE 1,8,Helv,,5.98,57.56,20.99,12.02,5,1342242816,128,OK,0,,8.00,5.58,18.29,19.69,12,1342177283,130,comment,0,,33.98,5.58,92.80,75.57,13,1350565888,130,Albert has already performed some preliminary experiments. Your first objective is to study the results of these experiments and suggest why he has not managed to produce any HGH so far.,0 &{ctrlID First objective ob2button init dlgInit setValue( ,"","") --DLL function retValue dialog(dlgBox getValue( ., "") buttonUp buttonUp dlgInit dlgBox dialog retValue |dlgInit ,b5,FALSE ,h12, ,s13,The behaviour of each promoter depends on the carbon source available. Your second objective is to determine the effect of the carbon source on the promoter and on HGH production. }dlgBox 524480,3,150,30,133,100,,,OBJECTIVE 2,8,Helv,,5.98,60.59,20.99,11.98,5,1342242816,128,OK,0,,8.00,8.57,18.29,19.69,12,1342177283,130,comment,0,,33.98,8.57,92.80,74.58,13,1350565888,130,The behaviour of each promoter depends on the carbon source available. Your second objective is to determine the effect of the carbon source on the promoter and on HGH production.,0 ctrlID Second objective ob3button init dlgInit setValue( ,"","") --DLL function retValue dialog(dlgBox getValue( ., "") buttonUp buttonUp dlgInit dlgBox dialog retValue dlgInit ,b5,FALSE ,h12, ,s13,The genotype of the strain affects the amount of product produced. Your third objective is to determine the effect that the choice of strain has on HGH production. dlgBox 524480,3,150,90,127,90,,,OBJECTIVE 3,8,Helv,,5.98,59.57,20.99,12.02,5,1342242816,128,OK,0,,8.00,7.59,18.29,19.69,12,1342177283,130,comment,0,,33.98,7.59,86.02,65.97,13,1350565888,130,The genotype of the strain affects the amount of product produced. Your third objective is to determine the effect that the choice of strain has on HGH production.,0 ctrlID Third objective ob4button init dlgInit setValue( ,"","") --DLL function retValue dialog(dlgBox getValue( ., "") buttonUp buttonUp dlgInit dlgBox dialog retValue dlgInit ,b5,FALSE ,h12, ,s13, Your final objective is simply to optimise HGH production for the promoters used. dlgBox 524480,3,150,130,127,90,,,OBJECTIVE 4,8,Helv,,5.98,59.57,20.99,12.02,5,1342242816,128,OK,0,,8.00,7.59,18.29,19.69,12,1342177283,130,comment,0,,35.31,6.77,86.02,65.97,13,1350565888,130, Your final objective is simply to optimise HGH production for the promoters used.,0 ctrlID Fourth objective "rustspeak" "albertspeak" "jeanspeak" enterPage leavePage enterPage rustspeak albertspeak jeanspeak leavePage !v# % *v- /.6 albertspeak Who are you? Oh, I remember, you're the new research officer. I applied for that job you know, but they turned me down. Said I was fine on the theory, but a bit long in the tooth for the practical side. Well, I can tell you that theory is important. You've got to think about what you're going to do before you run your experiment; plan it properly before you do it. When you've decided what your objectives are (Jean might be able to help you get some background information if you're stuck), go to the laboratory to plan and run your experiment. The data you generate will appear on the screen and will also be copied into your lab book. So if you want to look at your results at anytime, just click on the "LabBook" button when you've finished your current experiment. You can always return to the lab to do more experiments if necessary! I've already tried a few experiments and written the results at the beginning of the lab book. They didn't really work though, as no HGH was formed. I think Dr Rust wants you to explain this if you can! jeanspeak Hello, I'm Jean, the company librarian. Dr Rust has asked me to help you get settled in and give you some background information on your project. A lot of the screens you'll see during your project will have icons or buttons that you can point to for further information, or click on to make something happen. For example, point to the icons on the left or right-hand sides of this screen and I'll tell you more about them. You can use the buttons at the foot of every screen as navigation aids to find your way around. Just point at them and I'll tell you what they do, but only click on them when you actually want to go there! The title at the top of the screen will remind you where you are at present. As regards background information, you'll get some useful information in the library. Click on the library button below and go there now! Feel free to come back and see me anytime, by clicking on the Aims button... ob1hint Clicking anywhere on this area will show you the first objective you should aim to achieve. exithint Clicking the exit button will end your tutorial session. Only do this if you're sure you're finished as you will lose your results. aimshint Clicking the aims button will take you back to Dr Rust. He's quite a friendly chap really, and won't mind being disturbed!results. libhint Clicking the library button will take you to the library. Here you will find all sorts of useful background information which will help you with your project. You'd be well advised to pay a visit! labhint Clicking the laboratory button will take you to the lab where you can plan, prepare and run your experiments. Make sure you know what your objectives are before you start here.sed to pay a visit!sit!!! resultshint Clicking the LabBook button will let you examine your lab book where you have stored the results of all your experiments. Albert's preliminary experiments are also here.ere.re.sed to pay a visit!sit!!! qhint Clicking the report button will take you to the last section where your supervisor would like you to write him a report of the work you have done so far.ents are also here.e.re.sed to pay a visit!sit!!! ob2hint Clicking anywhere on this area will show you the second objective you should aim to achieve. ob3hint Clicking anywhere on this area will show you the third objective you should aim to achieve. rusthint Clicking anywhere on this area will allow Dr Rust to talk to you..st.should aim to achieve. jeanhint Clicking anywhere on this area is what allowed me to talk to you!ou should aim to achieve. titlehint This title reminds you what button you clicked to get to this screen. It is present on almost every screen. helphint Clicking the help button will give you general help on using the computer for MCB tutorials..ng this computer for MCB tutorials.ts. ob4hint Clicking anywhere on this area will show you the fourth objective you should aim to achieve. rustspeak WELCOME TO SCOTTISH BIOTECH. I hope you're going to be very happy working with us. We were very impressed with you at the interview and I have high hopes for you. I'm going to start you off working on our Hamster Growth Hormone (HGH) project. HGH is the latest idea in cattle feed supplements. Tests show that it vastly increases the animals' cheek capacity, thus improving their cud-chewing capabilities and significantly improving the efficiency of feed utilisation. The only disadvantage is that all the animals end up with stumpy tails and can't swat the flies away. What I want you to do is to find an appropriate HGH gene expression system which will maximise HGH production. To make this easier for you I've set four objectives. Click on the icons at the right-hand side of the screen and I'll explain them to you. There are two other people who you must speak to. Albert, your lab assistant, will tell you about some preliminary work he's done, and Jean, our librarian, will explain how to get all the information you will need to complete this work. Just click on their pictures at the left-hand side of the screen. Good luck!!!!l explain them to you. Good luck! alberthint Clicking anywhere on this area will allow Albert to talk to you....u should aim to achieve. fermenter mainbackground bkgd1 ackground tutorial --Move linkDLL statement handler "tbkdlg.dll" dialog( setValue( getValue( init dlgInit ,"","") function retValue dlgBox ", "") buttonUp buttonUp tbkdlg.dll dialog setValue getValue dlgInit dlgBox dialog retValue dlgInit ,b5,FALSE ,h12, ,s25,Yeast Foreign Gene Expression Systems Version 1.1 ,s27,A computer-based simulation exercise created by J.A.HAMNETT M.I.TAIT A.J.BROWN P.G.NEWMAN for the Department of Molecular and Cell Biology, University of Aberdeen, Scotland. ,s17,Electron micrograph reproduced from Baba and Baba, J.Cell Sci (1989) 94, 207 dlgBox 524480,5,70,50,223,154,,,ABOUT THIS TUTORIAL,8,Helv,,179.05,119.18,29.98,12.02,5,1342242816,128,OK,0,,14.78,10.63,18.29,19.69,12,1342177283,130,toolbook,0,,53.98,7.10,116.00,26.01,25,1350565889,130,Yeast Foreign Gene Expression Systems Version 1.1,0,,8.99,36.47,215.01,74.01,27,1342177280,130,A computer-based simulation exercise created by J.A.HAMNETT M.I.TAIT A.J.BROWN P.G.NEWMAN for the Department of Molecular and Cell Biology,0,,5.45,115.16,130.67,18.09,17,1350565889,130,Electron micrograph reproduced from Baba and Baba,0 ctrlID about this tutorial Laboratory yeast foreign gene expression systems yeast foreign gene expression systemsssss- -- - - - - - - yield ethanol Promoter: Strain: Substrate 1: Substrate 2: afterrrrrhanol yield: Ethanol yield::: Ethanol yield: Ethanol yield::::::l yield:tration: protease (A) MATa, ura3, pep4, prb1p4, prb14, prb1b1 promoter ADH2ose substrate1 Ethanolse substrate2 Ethanolse 8 hourss hours medium 0 unitstsl 0 mg/mlll00000004 g/l fillcolour 0,50,100 arrow arrow N & K SUBSTRATE 2 Arial mes New Roman Arial System Arial Arial Arial Times New Roman Arial Arial Arial Arial FOREIGN GENE EXPRESSION SYSTEMS MS Serif Arial Arial Arial Arial arial System Arial Times New Roman MS Serif Arial Arial System Symbol Arial System Arial Arial Wingdings Arial Arial Arial Arial Arial Arial Arial Arial Arial Arial System Arial Arial Times New Roman Symbol Symbol Times New Roman Arial Arial :PRINTLAYOUT arial :CONDITIONDATA Arial Times New Roman :REPORTDATA Arial Wingdings arial idNumber of this background = 1 arial Wingdings Arial report Arial Arial qxwtyEe xptno of page 4 of background labbkbackground text of field "checkprint" = "check" text of field "checkprint" = "check" J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$J$s Button Button steam Biomass Product 0 10 20 30 40 Time (hours)))rs)e (hours)hours)rs)urs)urs) - 0.66 - 1.66 - 2.66 0.4446 ln biomass (ln mg/ml)])) concn (Units/ml)4 Final HGH concn: Final ethanol concn:tration::::::::: yield 0 units ethanol 38 mg/ml labtxt RUNNING THE EXPERIMENT Use the yellow buttons on the right to plan, prepare and start an experiment. Your chosen parameters will appear on the grey bar above. Remember to keep an eye on the fermenter!fermentor display on the left. arrow arrow x P u SUBSTRATE 2 enterPage LeavePage leaveBook tbkdlg.dll eatclicks GlobalAlloc krnl386.exe GlobalAlloc GlobalLockPointer_ GlobalLock GlobalUnlock GlobalFree PeekMsg_ PeekMessage GlobalAlloc GlobalLockPointer_ GlobalFree PeekMsg_ PeekMsg_ PeekMsg_ GlobalUnlock GlobalFree krnl386.exe wFlags dwBytes LpMsg OnOrOff background joketxt "foreigngene" adhtxt galtxt pgktxt leavePage leavePage foreigngene foreigngene foreigngene foreigngene pgktxt galtxt adhtxt joketxt B 8"l Click on one of the topics on the left for further information. labbk background 4checkgraph /"labbk 400,566 - 11)*-10,0 60,87.625,100 "hideprint" "fly" prevents cursor changing arrow on entering ie overrides the handler enterBackground paste leaveBackground paste labbk background y5newPage checkgraph enterBackground pages pages leaveBackground hideprint -8-^- 0:0l1 5`7`7 hideprint pages 4checkgraph buttonUp buttonUp background checkgraph background button buttonUp buttonUp buttonUp background button Library LabBookENTTTT ExitProgram buttonUp buttonUp help button "helppage" buttonUp buttonUp helppage q button "questions1" buttonUp buttonUp questions1 buttonUp buttonUp buttonUp q button Report aims button "aims" buttonUp buttonUp target B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button results button "results1" buttonUp buttonUp results1 B"results buttonUp buttonUp buttonUp results button LabBook run button 4inoc "run1" buttonUp buttonUp buttonUp buttonUp buttonUp run button Laboratory Click arrows to turn pages of lab bookkkkkkkkkkk leftarrow "run1" "This /"labbk H10,0 eatclicks() buttonUp buttonUp This is the first page in the lab book pages labbk background eatclicks rightarrow "questions1" "This your lab /"labbk H-10,0 eatclicks() buttonUp buttonUp questions1 This is the last page of your lab book pages labbk background eatclicks buttonUp buttonUp buttonUp q button B"results buttonUp buttonUp buttonUp results button expdetails &(>'#( EXPERIMENT NO: Conditions Yeast strain: Promoter: Substrate1: Substrate2: Time of addition of substrate2: Final HGH concn: fly units/ml Final ethanol concn: :::::::::::::::::::::: flyconstant "fly" "bum" buttonUp buttonUp holdgraph 0 10 20 30 40 Time (hours) - 0.66 - 1.66 - 2.66 0.4446 ln biomass (ln mg/ml)])) V1.1S1 Biomass Product concn (Units/ml)4 flytxt Fly units! The number of flies on the cow's *@%!* is inversely proportional to the length of its tail!!ts tail!!!! "fly" "bum" buttonUp buttonUp Time (hours) results1 number Albert's no1 newgraph exptno (A)MATa, ura3 Ethanol Ethanol 2 hours 44mg/ml 0 fly units/ml flyconstant "fly" "bum" buttonUp buttonUp checkprint arrow arrow SUBSTRATE 2 studname reportbackground REPORTONSTTTT ExitProgram buttonUp buttonUp "helppage" buttonUp buttonUp helppage aims button "aims" buttonUp buttonUp B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button run button "medium" /"fermenter" "autoclave tape" inoc buttonUp buttonUp medium fermenter autoclave tape fermenter buttonUp buttonUp buttonUp run button Laboratory buttonUp buttonUp buttonUp run button background button buttonUp buttonUp background buttonUp buttonUp buttonUp background button Library buttonUp buttonUp buttonUp background button "questions1" buttonUp buttonUp questions1 buttonUp buttonUp buttonUp Report B"q" buttonUp buttonUp buttonUp results button "results1" buttonUp buttonUp results1 B"results buttonUp buttonUp buttonUp results button LabBook B"results buttonUp buttonUp buttonUp results button continue /!"^k buttonUp buttonUp Click here to continue studentname "0123456789" "0123456789" keyChar keyChar 0123456789 0123456789 questions1 You must now decide which experiments give you optimal results for each promoter. Enter your name and experiment numbers where indicated on the following page. You can get a copy of these experiments by clicking the "PRINT GRAPH" button. (Be patient, it can take a while!) When you have done this, you have completed the experimental part of this tutorial, but make sure you have made all the notes you require, as your results will not be saved when you exit the tutorial. You should now open the file "YEAST.W" in Microsoft Works and complete the report/questions you will find there. Remember to put your name and your partner's name at the top of the page! Hand the completed report (graphs and questions/answers) to your tutor by the date indicated in your handout.eted report (graphs and questions/answers) to your tutor by the date indicated in your handout.ns/answers) to your tutor.... enterpage enterpage continue mainbackground buttonup buttonup ExitProgram quit buttonUp buttonUp help button "helppage" buttonUp buttonUp helppage aims button "aims" buttonUp buttonUp target B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button results button "results1" buttonUp buttonUp results1 B"results buttonUp buttonUp buttonUp results button LabBook run button "run1" buttonUp buttonUp buttonUp buttonUp buttonUp run button Laboratory buttonUp buttonUp background background button buttonUp buttonUp buttonUp background button Library "questions1" buttonUp buttonUp questions1 Report buttonUp buttonUp buttonUp q button B"results buttonUp buttonUp buttonUp results button questions1 diauxicgrowth results1 start1 foreigngene questions2 newgraph (A) MATa, ura3 GAL1) Ethanol Ethanol 2 hours hours 0 fly units/ml flyconstant "fly" "bum" buttonUp buttonUp 44 mg/mlS Albert's no 2 arrow arrow SUBSTRATE 2 studname checkprint 4checkgraph, nosave ZmyList "exp1" "exp2" "exp3" "Remember type a EACH box." "You have xperformed many experiments!" "Experiment 0!?! ygotta be kiddin'!." "studentname" your xsaved one these 8." & \ "Are sure want Y graphs " && ( &&","&& ( ) &&" "&& ( ) &&"?" f"Print" "printing" you don't printed /"labbk "hideprint" --match the chosen correct labbook /" + 1 "checkprint" "studname" "(" & & ") Name:" && 1000,1000,1000,1000 " = "" "bar" 3195,4560,3250,4755 3195,4560,3250+a,4755 Za=2200 B"printgraphbut" 3195,4560,3250,4755 "questions2" eatclicks() buttonUp buttonUp continue Remember to type a number into EACH box. You have not performed this many experiments! Experiment 0!?! You gotta be kiddin'!. studentname Remember to type in your name! studentname You have not saved one of these experiments in your lab book. Go back and check. Are you sure you want to print the graphs from experiments Print printing labbk background 0,100,0 hideprint labbk background labbk background checkprint check studname studname ) Name: studentname checkprint checkprint text of field "checkprint" = "check" labbk background labbk background checkprint studname studname studname printing printgraphbut printing buttonup continue studentname questions2 questions2 questions2 questions2 eatclicks myList checkgraph nosave Times New Roman Arial Times New Roman "System 9|acgr iV{3o wYPv`gb LrdUkE ~Uxl|rcwp zuic~a umPsUz szqizY ^w`yx ed_}hq oz]rA lqWi[T ]ns=Y Whi^Ox zr~iW S^^OM WqbGb^ FIVv[ v{Kjfx Fn{G`W Zow*` Wj^}i OwGxb 5ZVkG?kIm \epBb KV\agWgXOu Hb[:\ Eobdvm @Q3pvx e7ucX 4:H{pc{ 8Cz3M}Dj fgobq Z|N[A =}kb^ >SSwd 0zE>q <TDRvk XXhQl HFo~^ HbmNR uK;vdK]@1R Jb(<7/ nNKivzYG l2uwac ]hCXk]i N`eYq^ cBg|I\ ?dwD>^KR 'otkH ry\P9 Q{P`FS IHg_BJEI +m[[< 6K[Ov` yB_uo \49iW9@Igv ~~u7{ne BB1<HEwMS c~anao s5McpW msOULVWs VLp@r- bR~zC "XqQc Mavtk ]@E\/QT] {pbhu @xInmO ,WH[0 DXXo\ aSs~W hzqkE ^;f97 Zd>;b\ P.t|cGQ [[y0@s G?H[}IdDEQ tiWpU| ^Scm~4 LthRNJr_ 8c7]f rf]4IG; ~{cuo \UX9WAeJwJ BhXYjKMQ<L _lGJN WwyHS %)Z_hY|h vb}IZa Xu`W5 ^}}`\# i[d\QR wi6;H;MQO 9jvz\ DL^w[ =NGSKKRsKV `~~iKk|< ilPvG ekw+q sExHR ga[ub @vfG5w; xsCai |b9R_hzB >|R\` ip]MVn^k^ q0dp{ py>xub uV^`f MzgOR S~Qar {ANVx UGfjw As[{q ^adro FjAg3> ga4zx K|4{_Oe 6sMGU i~R]yP XT}~M}RN euk*;uw [Mf[[~ WiKfX yW|T@U ]PgO[ ^|sP_ p^Q{\ MJrEj c|Sy`Ya P8ozg{^ N{Ak| V+Pb~ m\ojS gRqU7 N7rcs aoe`} Y|iq\ XIaUliK\`^ HM}FaQ UV9R` 6qbND`h #wnzsK ivGgw e\aYXm`?So \\8Iga OmQLZ MCipy} wnaOY 5lLaLMu 0k:dr >S}z[c X>sw*[ gv\4J itY}8 IiK_G`: nVdMoCz =vgQK `w`QxW MYnaV ej.`u |K[Xbe m>vSX ?HdSv pK}U~ ucHhlk uYct| }mu(`ITm ^VM^Og`F cMX+^ JxGci K`dd9db \=bmT Rkc-sh --sets up path multimedia toolbk c"tbkmm.sbk" "e:\toolbook\ -- prompt --remember change final version [ will suspend execution the event error DECLARE AND INITIALISE SYSTEM VARIABLES 4inoc, checkgraph,substrate1,substrate2,promoter,tadd,protease,planinit, ferment, checkalbres 4checkalbert, checkjean,objective1, objective2, objective3, objective4, pagecounter 4culturecolor, bud, budding, begintime, finishtime, looptime, checkplan, checkprepare 4checksterilise,findpage,mouth,nosave used %a flag buttons have been pressed -- correct order - see incremented Hone each drawn on "start1" asure you library\ 4results ( aims Lalbertspeak read ( jeanspeak etc are %flags objectives laboratory section initialise yeast on title graphs saved labbook contain a list expts which Al's expression on usedbefore student has program --following values required drawing /"fermenter" "Ethanol" "ADH2" "8 hours" "(A)MATa,ura3" SET UP MENU c"File" c"Text" c"Page" c"Help" c"&Go e"&Title c"Go c"Go e"&Aims" c"Go e"&Library" c"Go e"Laborator&y" c"Go e"La&bBook" c"Go e"Re&port" c"Go "Ab&out tutorial" c"help" "&Quit" WORK OUT SPEED OF COMPUTER BY TIMING A LOOP cx<5000 speed handler ( Reader "This created Department Molecular Cell Biology, University Aberdeen, Scotland. It freely distributable among educational establishments xaltered adapted way without G permission J.A.Hamnett, 1993" --Move linkDLL statement "tbkdlg.dll" dialog( setvalue( getvalue( "yield" "ethanol" "labtxt" "rustspeak" "medium" "autoclave tape" "Have f"Yes" want available introductory information?"\ "yes" Aboutthistutorial /"bkgd1" Gotohelppage TitlePage "startpage" Laboratory LabBook "results1" Report "questions1" "You xcompleted enough experiments. Do really quit?"\ "&No" SysSuspendMessages LeavePage unlinkDLL " TOOLBOX FOR TOOLBOOK Redmon Group ,Tool: EatClicks --Original Programming H: Claude Osteen Asymetrix --Modified Jeffrey S. Howard --Designs H: John Veronica Cruz Daniel Lazenby --Last Date : October 22, 1992 acknowledges Technical Support Staff some contained --The original eatClicks developed --was modified work fWindows 3.1 -- Function: Eatclicks -- Purpose: Cancel pending keyboard mouse messages -- Parameters: None -- Returns: 0 positive sucessful eatclicks OnOrOff Local wFlags, dwBytes, cnt, hMsg, LpMsg -- Link windows functions allocates a block memory only xdone already. 0 -- GlobalAlloc 32 -- MSG data structure, froom spare. Linkdll "krnl386.exe" WORD , DWORD) POINTER GlobalLockPointer_ = 9GlobalUnlock( NGlobalFree( X PeekMsg_ = PeekMessage( = 0 -- Call unwanted -- Mouse , 512, 521, 1) <> 0 -- Keyboard presses , 256, 264, 1) <> 0 -- Menu Accelerators , 111, 112, 1) <> 0 unLinkdll " istutorial Library enterBook Laboratory LabBook Gotohelppage Report TitlePage enterPage Aboutthistutorial LeavePage leaveBook eatclicks enterBook tbkmm.sbk e:\toolbook\tbkmm.sbk substrate1 fermenter Ethanol substrate2 fermenter Ethanol promoter fermenter fermenter 8 hours protease fermenter (A)MATa,ura3 &Go to &Title page Go to &Help Go to &Aims Go to &Library Go to Laborator&y Go to La&bBook Go to Re&port Go to &Help Ab&out this tutorial Go to &help page &Quit clear paste seconds sizetopage This program has been created for the Department of Molecular and Cell Biology, University of Aberdeen, Scotland. It is freely distributable among educational establishments in the UK as long as it is not altered or adapted in any way without the permission of the author. J.A.Hamnett, 1993 tbkdlg.dll dialog setvalue getvalue yield ethanol labtxt albertspeak jeanspeak rustspeak medium fermenter autoclave tape fermenter Have you used this tutorial before? Do you want to read all the available introductory information? usedbefore checksterilise findpage mouth nosave culturecolor budding begintime finishtime looptime checkplan checkprepare checkalbert checkjean objective1 objective2 objective3 objective4 pagecounter checkgraph substrate1 substrate2 promoter protease planinit ferment checkalbres Aboutthistutorial buttonUp tutorial bkgd1 Gotohelppage helppage TitlePage startpage helppage Library background Laboratory LabBook results1 Report questions1 You have not completed enough experiments. Do you really want to quit? This will end the tutorial. Are you sure you want to quit? checkgraph enterPage LeavePage leaveBook tbkdlg.dll eatclicks GlobalAlloc krnl386.exe GlobalAlloc GlobalLockPointer_ GlobalLock GlobalUnlock GlobalFree PeekMsg_ PeekMessage GlobalAlloc GlobalLockPointer_ GlobalFree PeekMsg_ PeekMsg_ PeekMsg_ GlobalUnlock GlobalFree krnl386.exe wFlags dwBytes LpMsg OnOrOff fermenter " ,d0F2v2 `X`lb ExitProgram buttonUp buttonUp help button "helppage" buttonUp buttonUp helppage q button "questions1" buttonUp buttonUp questions1 buttonUp buttonUp buttonUp q button Report aims button "aims" buttonUp buttonUp target B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button B"aims buttonUp buttonUp buttonUp aims button results button 4findpage checkgraph=0 buttonUp buttonUp checkgraph findpage B"results buttonUp buttonUp buttonUp results button LabBook LABORATORYYTTT Plan experiment 4checkplan B"dialog" "You have now planned your experiment! The values /chosen are displayed the grey bar screen." eatclicks() buttonUp buttonUp buttonUp dialog You have now planned your first experiment! The values you have chosen are displayed in the grey bar at the top of the screen. eatclicks checkplan o c$? o /"? !- " Prepare experiment prepare 4checkprepare medium 60,90,100 "You've already prepared the E. Albert xamused!" "Your has been added efermenter vessel." eatclicks() buttonUp buttonUp medium You've already prepared the medium. Albert is not amused! medium Your medium has been prepared and added to the fermenter vessel. eatclicks medium checkprepare |#u!k Sterilise sterilise 4looptime, checksterilise "medium" "Hadn't you better prepare the V everything?" eatclicks() "autoclave tape" "You've already sterilised zfermenter once!" "steam" "newgraph" buttonUp buttonUp medium Hadn't you better prepare the medium before you sterilise everything? eatclicks autoclave tape You've already sterilised the fermenter once! autoclave tape steam steam You've now sterilised the fermenter. newgraph Wungroup eatclicks looptime checksterilise Inoculate inoc2 buttonUp buttonUp background background button buttonUp buttonUp buttonUp background button Library pH electrode v=N=s= |>T>y> speed &9~@, speedknob rbaffle rbaffle "bubble" "components" "stirrer" mouseEnter mouseLeave mouseEnter bubble components stirrer components mouseLeave components components right centre "bar" "centre" buttonup buttonup centre centre right right centre centre right right ZK2KWK run button 4inoc "medium" "autoclave tape" buttonUp buttonUp medium autoclave tape buttonUp buttonUp buttonUp run button buttonUp buttonUp buttonUp run button Laboratory Strain: Promoter: Substrate 1: Substrate 2: after medium PRFRcolor dRfillcolour 0,50,100 autoclave tape protease c@SeS (A)MATa,ura33, pep4, prb11 promoter ADH2ose substrate1 rTJToT Ethanolse substrate2 Ethanolse xUPUuU 8 hourss hours Dialog UctrlID UdlgBox 524480,22,26,24,307,192,,,Plan experiment,8,Helv,,7.62,9.68,18.29,19.69,63,1342177283,130,toolbook,0,promoter,33.41,11.16,62.10,54.28,16,1342177287,128, Promoter,0,strain,106.40,10.95,140.15,54.28,29,1342177287,128, Yeast strain,0,substrate1,33.60,75.49,61.87,54.28,33,1342177287,128, Substrate 1,0,substrate2,106.40,75.49,140.15,54.28,37,1342177287,128, Substrate 2 ,0,addedafter,175.20,88.74,44.80,11.45,51,1342177280,130,Added after:,0,,7.62,136.33,293.07,36.18,182,1342177280,130,_________________________________________________________________________ Plan your experiment by clicking on your choice of promoter,0,ok,255.73,14.56,45.33,16.33,12,1342242817,128,&OK,0,cancel,255.85,40.90,44.80,16.08,14,1342242816,128,&Cancel,0,adh,40.80,23.84,42.90,10.05,21,1342308361,128,ADH2,0,gal,40.80,35.90,41.07,10.05,22,1342177289,128,GAL1,0,pgk,40.80,48.98,41.07,10.05,171,1342177289,128,PGK1,0,straina,114.25,23.84,125.10,10.46,40,1342308361,128,Strain A: MATa ura3,0,strainb,114.25,36.64,123.62,10.46,39,1342177289,128,Strain B: MATa ura3 pep4 prb1,0,strainc,114.25,49.64,125.10,10.46,38,1342177289,128,Strain C: MATa ura3 pep4,0,ethanol,41.22,89.15,50.90,10.75,41,1342308361,128,Ethanol,0,galactose,40.80,101.78,50.90,10.75,43,1342177289,128,Galactose,0,glucose,41.03,114.58,50.93,10.75,42,1342177289,128,Glucose,0,ethanol2,114.25,89.15,50.93,11.16,46,1342308361,128,Ethanol,0,galactose2,114.25,102.73,50.93,11.12,44,1342177289,128,Galactose,0,glucose2,114.25,115.57,50.93,11.16,45,1342177289,128,Glucose,0,tadd,174.63,100.80,59.43,54.31,227,1352859715,133,,0 \dlgInit ,h63, groupbox promoter,g16, Promoter groupbox strain,g29, Yeast strain groupbox substrate1,g33, Substrate 1 groupbox substrate2,g37, Substrate 2 static addedafter,s51,Added after: ,s182,_________________________________________________________________________ Plan your experiment by clicking on your choice of promoter, yeast strain, substrates and the time you want to add the second substrate, then click OK to save these settings. button ok,b12,TRUE button cancel,b14,FALSE button adh,b21,TRUE button gal,b22,FALSE button pgk,b171,FALSE button straina,b40,TRUE button strainb,b39,FALSE button strainc,b38,FALSE button ethanol,b41,TRUE button galactose,b43,FALSE button glucose,b42,FALSE button ethanol2,b46,TRUE button galactose2,b44,FALSE button glucose2,b45,FALSE combobox tadd,c227,2 hours 4 hours 6 hours &8 hours 10 hours 12 hours 14 hours 16 hours 18 hours 20 hours 22 hours 24 hours 26 hours 28 hours 30 hours 32 hours 34 hours 36 hours 38 hours 40 hours Plan experiment inoculate Inoculate buttonUp buttonUp buttonUp q button B"results buttonUp buttonUp buttonUp results button --sets up path multimedia toolbk c"tbkmm.sbk" "This program needs runtime Pook work properly. If you have installed, asure the [Toolbook] section your win.ini file Frestart sysSuspendMessages "e:\toolbook\ -- prompt --remember change final version ] will suspend execution event error DECLARE AND INITIALISE SYSTEM VARIABLES 4inoc, checkgraph,substrate1,substrate2,promoter,tadd,protease,planinit, ferment, checkalbres 4checkalbert, checkjean,objective1, objective2, objective3, objective4, pagecounter 4culturecolor, bud, budding, begintime, finishtime, looptime, checkplan, checkprepare 4checksterilise,findpage,mouth,nosave used %a flag buttons been pressed -- correct order - see incremented Hone each drawn on "start1" library\ 4results ( aims albertspeak read ( jeanspeak etc are %flags objectives laboratory initialise yeast on title graphs saved labbook contain a list expts which Al's expression on usedbefore student has --following values required drawing /"fermenter" "Ethanol" "ADH2" "8 hours" "(A)MATa,ura3" SET UP MENU c"File" c"Text" c"Page" c"Help" c"&Go e"&Title c"Go c"Go e"&Aims" c"Go e"&Library" c"Go e"Laborator&y" c"Go e"La&bBook" c"Go e"Re&port" c"Go "Ab&out tutorial" c"help" "&Quit" WORK OUT SPEED OF COMPUTER BY TIMING A LOOP cx<5000 speed handler ( Reader created x Department Molecular Cell Biology, University Aberdeen, Scotland. It freely distributable among educational establishments xaltered adapted way without permission J.A.Hamnett, 1993" --Move linkDLL statement "tbkdlg.dll" dialog( setvalue( getvalue( "yield" "ethanol" "labtxt" "rustspeak" "medium" "autoclave tape" "Have f"Yes" want available introductory information?"\ "yes" Aboutthistutorial /"bkgd1" Gotohelppage TitlePage "startpage" Laboratory LabBook "results1" Report "questions1" "You xcompleted enough experiments. Do really quit?"\ "&No" SysSuspendMessages LeavePage unlinkDLL " TOOLBOX FOR TOOLBOOK Redmon Group : EatClicks --Original Programming H: Claude Osteen Asymetrix --Modified Jeffrey S. Howard --Designs H: John Veronica Cruz Daniel Lazenby --Last Date : October 22, 1992 acknowledges Technical Support Staff some contained --The original eatClicks developed --was modified fWindows 3.1 -- Function: Eatclicks -- Purpose: Cancel pending keyboard mouse messages -- Parameters: None -- Returns: 0 positive sucessful eatclicks OnOrOff Local wFlags, dwBytes, cnt, hMsg, LpMsg -- Link windows functions allocates a block memory only xdone already. 0 -- GlobalAlloc 32 -- MSG data structure, froom spare. Linkdll "krnl386.exe" WORD , DWORD) POINTER GlobalLockPointer_ = 9GlobalUnlock( NGlobalFree( X PeekMsg_ = PeekMessage( = 0 -- Call unwanted -- Mouse , 512, 521, 1) <> 0 -- Keyboard presses , 256, 264, 1) <> 0 -- Menu Accelerators , 111, 112, 1) <> 0 unLinkdll " istutorial Library enterBook Laboratory LabBook Gotohelppage Report TitlePage enterPage Aboutthistutorial LeavePage leaveBook eatclicks enterBook tbkmm.sbk This program needs runtime multimedia toolbook to work properly. If you have this installed, make sure the [Toolbook] section of your win.ini file contains the line 'startupSysBooks=tbkmm.sbk', then restart the program. substrate1 fermenter Ethanol substrate2 fermenter Ethanol promoter fermenter fermenter 8 hours protease fermenter (A)MATa,ura3 &Go to &Title page Go to &Help Go to &Aims Go to &Library Go to Laborator&y Go to La&bBook Go to Re&port Go to &Help Ab&out this tutorial Go to &help page &Quit clear paste seconds sizetopage This program has been created for the Department of Molecular and Cell Biology, University of Aberdeen, Scotland. It is freely distributable among educational establishments in the UK as long as it is not altered or adapted in any way without the permission of the author. J.A.Hamnett, 1993 tbkdlg.dll dialog setvalue getvalue yield ethanol labtxt albertspeak jeanspeak rustspeak medium fermenter autoclave tape fermenter Have you used this tutorial before? Do you want to read all the available introductory information? usedbefore checksterilise findpage mouth nosave culturecolor budding begintime finishtime looptime checkplan checkprepare checkalbert checkjean objective1 objective2 objective3 objective4 pagecounter checkgraph substrate1 substrate2 promoter protease planinit ferment checkalbres W={ Aboutthistutorial buttonUp tutorial bkgd1 Gotohelppage helppage TitlePage startpage helppage Library background Laboratory LabBook results1 Report questions1 You have not completed enough experiments. Do you really want to quit? This will end the tutorial. Are you sure you want to quit? checkgraph enterPage LeavePage leaveBook tbkdlg.dll eatclicks GlobalAlloc krnl386.exe GlobalAlloc GlobalLockPointer_ GlobalLock GlobalUnlock GlobalFree PeekMsg_ PeekMessage GlobalAlloc GlobalLockPointer_ GlobalFree PeekMsg_ PeekMsg_ PeekMsg_ GlobalUnlock GlobalFree krnl386.exe wFlags dwBytes LpMsg OnOrOff multimedia toolbk c"tbkmm.sbk" "This program needs runtime Pook work properly. If you have installed, asure the [Toolbook] section your win.ini file Frestart "e:\toolbook\ sysSuspendMessages -- prompt --remember change final version ~ will suspend execution event error DECLARE AND INITIALISE SYSTEM VARIABLES 4inoc, checkgraph,substrate1,substrate2,promoter,tadd,protease,planinit, ferment, checkalbres 4checkalbert, checkjean,objective1, objective2, objective3, objective4, pagecounter 4culturecolor, bud, budding, begintime, finishtime, looptime, checkplan, checkprepare 4checksterilise,findpage,mouth,nosave used %a flag buttons been pressed -- correct order - see incremented Hone each drawn on "start1" library\ 4results ( aims albertspeak read ( jeanspeak etc are %flags objectives laboratory initialise yeast on title graphs saved labbook contain a list expts which Al's expression on usedbefore student has --following values required drawing /"fermenter" "Ethanol" "ADH2" "8 hours" "(A)MATa,ura3" SET UP MENU c"File" c"Text" c"Page" c"Help" c"&Go e"&Title c"Go c"Go e"&Aims" c"Go e"&Library" c"Go e"Laborator&y" c"Go e"La&bBook" c"Go e"Re&port" c"Go "Ab&out tutorial" c"help" "&Quit" WORK OUT SPEED OF COMPUTER BY TIMING A LOOP cx<5000 speed handler ( Reader created q Department Molecular Cell Biology, University Aberdeen, Scotland. It freely distributable among educational establishments xaltered adapted way without permission J.A.Hamnett, 1993" --Move linkDLL statement "tbkdlg.dll" dialog( setvalue( getvalue( "yield" "ethanol" "labtxt" "rustspeak" "medium" "autoclave tape" "Have f"Yes" want available introductory information?"\ "yes" Aboutthistutorial /"bkgd1" Gotohelppage TitlePage "startpage" Laboratory LabBook "results1" Report "questions1" "You xcompleted enough experiments. Do really quit?"\ "&No" SysSuspendMessages LeavePage -- unlinkDLL " TOOLBOX FOR TOOLBOOK Redmon Group : EatClicks --Original Programming H: Claude Osteen Asymetrix --Modified Jeffrey S. Howard --Designs H: John Veronica Cruz Daniel Lazenby --Last Date : October 22, 1992 acknowledges Technical Support Staff some contained --The original eatClicks developed --was modified fWindows 3.1 -- Function: Eatclicks -- Purpose: Cancel pending keyboard mouse messages -- Parameters: None -- Returns: 0 positive sucessful eatclicks OnOrOff Local wFlags, dwBytes, cnt, hMsg, LpMsg -- Link windows functions allocates a block memory only xdone already. 0 -- GlobalAlloc 32 -- MSG data structure, froom spare. Linkdll "krnl386.exe" WORD , DWORD) POINTER GlobalLockPointer_ = 9GlobalUnlock( NGlobalFree( X PeekMsg_ = PeekMessage( = 0 -- Call unwanted -- Mouse , 512, 521, 1) <> 0 -- Keyboard presses , 256, 264, 1) <> 0 -- Menu Accelerators , 111, 112, 1) <> 0 unLinkdll " istutorial Library enterBook Laboratory LabBook Gotohelppage Report TitlePage enterPage Aboutthistutorial LeavePage leaveBook eatclicks enterBook tbkmm.sbk This program needs runtime multimedia toolbook to work properly. If you have this installed, make sure the [Toolbook] section of your win.ini file contains the line 'startupSysBooks=tbkmm.sbk', then restart the program. substrate1 fermenter Ethanol substrate2 fermenter Ethanol promoter fermenter fermenter 8 hours protease fermenter (A)MATa,ura3 &Go to &Title page Go to &Help Go to &Aims Go to &Library Go to Laborator&y Go to La&bBook Go to Re&port Go to &Help Ab&out this tutorial Go to &help page &Quit clear paste seconds sizetopage This program has been created for the Department of Molecular and Cell Biology, University of Aberdeen, Scotland. It is freely distributable among educational establishments in the UK as long as it is not altered or adapted in any way without the permission of the author. J.A.Hamnett, 1993 tbkdlg.dll dialog setvalue getvalue yield ethanol labtxt albertspeak jeanspeak rustspeak medium fermenter autoclave tape fermenter Have you used this tutorial before? Do you want to read all the available introductory information? usedbefore checksterilise findpage mouth nosave culturecolor budding begintime finishtime looptime checkplan checkprepare checkalbert checkjean objective1 objective2 objective3 objective4 pagecounter checkgraph substrate1 substrate2 promoter protease planinit ferment checkalbres W={ Aboutthistutorial buttonUp tutorial bkgd1 Gotohelppage helppage TitlePage startpage helppage Library background Laboratory LabBook results1 Report questions1 You have not completed enough experiments. Do you really want to quit? This will end the tutorial. Are you sure you want to quit? checkgraph enterPage LeavePage leaveBook eatclicks GlobalAlloc krnl386.exe GlobalAlloc GlobalLockPointer_ GlobalLock GlobalUnlock GlobalFree PeekMsg_ PeekMessage GlobalAlloc GlobalLockPointer_ GlobalFree PeekMsg_ PeekMsg_ PeekMsg_ GlobalUnlock GlobalFree krnl386.exe wFlags dwBytes LpMsg OnOrOff --sets up path multimedia toolbk c"tbkmm.sbk" "This program needs runtime Pook work properly. If you have installed, asure the [Toolbook] section your win.ini file Frestart sysSuspendMessages "e:\toolbook\ -- prompt --remember change final version ] will suspend execution event error DECLARE AND INITIALISE SYSTEM VARIABLES 4inoc, checkgraph,substrate1,substrate2,promoter,tadd,protease,planinit, ferment, checkalbres 4checkalbert, checkjean,objective1, objective2, objective3, objective4, pagecounter 4culturecolor, bud, budding, begintime, finishtime, looptime, checkplan, checkprepare 4checksterilise,findpage,mouth,nosave used %a flag buttons been pressed -- correct order - see incremented Hone each drawn on "start1" library\ 4results ( aims albertspeak read ( jeanspeak etc are %flags objectives laboratory initialise yeast on title graphs saved labbook contain a list expts which Al's expression on usedbefore student has --following values required drawing /"fermenter" "Ethanol" "ADH2" "8 hours" "(A)MATa,ura3" SET UP MENU c"File" c"Text" c"Page" c"Help" c"&Go e"&Title c"Go c"Go e"&Aims" c"Go e"&Library" c"Go e"Laborator&y" c"Go e"La&bBook" c"Go e"Re&port" c"Go "Ab&out tutorial" c"help" "&Quit" WORK OUT SPEED OF COMPUTER BY TIMING A LOOP cx<5000 speed handler ( Reader created x Department Molecular Cell Biology, University Aberdeen, Scotland. It freely distributable among educational establishments xaltered adapted way without permission J.A.Hamnett, 1993" --Move linkDLL statement "tbkdlg.dll" dialog( setvalue( getvalue( "yield" "ethanol" "labtxt" "rustspeak" "medium" "autoclave tape" "Have f"Yes" want available introductory information?"\ "yes" Aboutthistutorial /"bkgd1" Gotohelppage TitlePage "startpage" Laboratory LabBook "results1" Report "questions1" "You xcompleted enough experiments. Do really quit?"\ "&No" SysSuspendMessages LeavePage -- unlinkDLL " TOOLBOX FOR TOOLBOOK Redmon Group : EatClicks --Original Programming H: Claude Osteen Asymetrix --Modified Jeffrey S. Howard --Designs H: John Veronica Cruz Daniel Lazenby --Last Date : October 22, 1992 acknowledges Technical Support Staff some contained --The original eatClicks developed --was modified fWindows 3.1 -- Function: Eatclicks -- Purpose: Cancel pending keyboard mouse messages -- Parameters: None -- Returns: 0 positive sucessful eatclicks OnOrOff Local wFlags, dwBytes, cnt, hMsg, LpMsg -- Link windows functions allocates a block memory only xdone already. 0 -- GlobalAlloc 32 -- MSG data structure, froom spare. Linkdll "krnl386.exe" WORD , DWORD) POINTER GlobalLockPointer_ = 9GlobalUnlock( NGlobalFree( X PeekMsg_ = PeekMessage( = 0 -- Call unwanted -- Mouse , 512, 521, 1) <> 0 -- Keyboard presses , 256, 264, 1) <> 0 -- Menu Accelerators , 111, 112, 1) <> 0 unLinkdll " istutorial Library enterBook Laboratory LabBook Gotohelppage Report TitlePage enterPage Aboutthistutorial LeavePage leaveBook eatclicks enterBook tbkmm.sbk This program needs runtime multimedia toolbook to work properly. If you have this installed, make sure the [Toolbook] section of your win.ini file contains the line 'startupSysBooks=tbkmm.sbk', then restart the program. substrate1 fermenter Ethanol substrate2 fermenter Ethanol promoter fermenter fermenter 8 hours protease fermenter (A)MATa,ura3 &Go to &Title page Go to &Help Go to &Aims Go to &Library Go to Laborator&y Go to La&bBook Go to Re&port Go to &Help Ab&out this tutorial Go to &help page &Quit clear paste seconds sizetopage This program has been created for the Department of Molecular and Cell Biology, University of Aberdeen, Scotland. It is freely distributable among educational establishments in the UK as long as it is not altered or adapted in any way without the permission of the author. J.A.Hamnett, 1993 tbkdlg.dll dialog setvalue getvalue yield ethanol labtxt albertspeak jeanspeak rustspeak medium fermenter autoclave tape fermenter Have you used this tutorial before? Do you want to read all the available introductory information? usedbefore checksterilise findpage mouth nosave culturecolor budding begintime finishtime looptime checkplan checkprepare checkalbert checkjean objective1 objective2 objective3 objective4 pagecounter checkgraph substrate1 substrate2 promoter protease planinit ferment checkalbres W={ Aboutthistutorial buttonUp tutorial bkgd1 Gotohelppage helppage TitlePage startpage helppage Library background Laboratory LabBook results1 Report questions1 You have not completed enough experiments. Do you really want to quit? This will end the tutorial. Are you sure you want to quit? checkgraph enterPage LeavePage leaveBook eatclicks GlobalAlloc krnl386.exe GlobalAlloc GlobalLockPointer_ GlobalLock GlobalUnlock GlobalFree PeekMsg_ PeekMessage GlobalAlloc GlobalLockPointer_ GlobalFree PeekMsg_ PeekMsg_ PeekMsg_ GlobalUnlock GlobalFree krnl386.exe wFlags dwBytes LpMsg OnOrOff