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M9480878.TXT
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1994-09-05
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Document 0878
DOCN M9480878
TI Stabilization and expression of HIVMN genes in Salmonella typhimurium.
DT 9410
AU Sizemore D; Branstrom A; Sadoff J; Warren R; Walter Reed Army Inst. of
Res., Washington, DC.
SO Abstr Gen Meet Am Soc Microbiol. 1994;94:151 (abstract no. E-46). Unique
Identifier : AIDSLINE ASM94/94313067
AB Stabilization and expression of recombinant plasmids in Salmonella
carriers for oral vaccination has proven problematic. Previously, we
showed that pSC101 derived plasmids were maintained in Salmonella
carriers without selection. However, foreign genes were expressed at low
levels. We report the construction of a novel plasmid vector that
contains origins of replication from pSC101 and pMB1. This vector has a
higher copy number and increased expression when compared with the
pSC101 vector. Regions of the gag, env, vif, pol and nef genes from the
HIVMN genome were PCR amplified and cloned into pAB102 and placed under
the control of plac. An asd- mutant of the S. typhimurium strain WS1321,
which lacks the large virulence plasmid was used as a carrier of the
recombinant plasmids. In vivo stability of plasmids pAB102, pAB102::gag
and pAB102::vif was examined over a 28-day period in BALB/c mice given
an oral dose of 4-6 x 10(9) bacteria. Increasing numbers of each
construct were cultured from the spleens of mice on days 3, 7 and 10.
Only 2 mice were culture positive by day 28. Immune responses were
measured in mice infected with pAB102::gag. No CTL activity or serum
antibody was detected above controls. The pagC promoter, which is known
to be active in macrophages, is currently being evaluated to increase in
vivo expression.
DE Animal Cloning, Molecular/*METHODS Genes, gag *Genes, Viral Genome,
Viral HIV/GROWTH & DEVELOPMENT/*GENETICS HIV
Antibodies/ANALYSIS/BIOSYNTHESIS Mice Mice, Inbred BALB C Plasmids
Polymerase Chain Reaction/METHODS Promoter Regions (Genetics)
Salmonella typhimurium/*METABOLISM/PATHOGENICITY Virulence MEETING
ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).