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M94A3314.TXT
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1994-10-25
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Document 3314
DOCN M94A3314
TI HIV LTR drives gene expression in mouse and human eggs.
DT 9412
AU Kiessling A; Crowell R; Michalopoulos J; Waldorf T; Deaconess/Harvard
Surgical Service, Faulkner Hospital, Boston, MA; 02130.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):104 (abstract no. PA0034). Unique
Identifier : AIDSLINE ICA10/94369261
AB OBJECTIVE: To determine if fertilized eggs would express HIV proteins.
METHODS: Mouse zygotes and human polypronuclear eggs were microinjected
with DNA constructs containing HIV-LTR linked to lacZ reporter gene.
Mouse zygotes were microinjected with 2 to 10,000 copies of plasmid
pHIV-LTR, cultured for 48 hours, fixed in paraformaldehyde and stained
for expression of beta-galactosidase. A plasmid construct containing
lacZ without promoter was microinjected as control. Four polypronuclear
human eggs were microinjected with approximately 6,000 copies of HIV-LTR
reporter gene construct, and two polypronuclear human eggs were
microinjected with control. The human eggs were also cultured for 48
hours and then stained for beta-galactosidase. RESULTS: Twenty five of
the 111 mouse zygotes microinjected with pHIV-LTR were positive for
beta-galactosidase. In contrast, all four of the human polypronuclear
eggs microinjected with the HIV-LTR reporter gene stained highly
positive for lacZ expression. None of the 22 zygotes nor the two
polypronuclear human eggs microinjected with control construct exhibited
staining. DISCUSSION AND CONCLUSIONS: These results indicate that the
HIV-LTR promoter is active in mouse zygotes, in contrast to earlier
reports that exogenous retroviruses are silenced in early embryos, but
in keeping with recent reports that the LTR of Rous sarcoma virus is
expressed in early embryos. Importantly, the HIV-LTR reporter gene
appears to be readily expressed in polypronuclear human eggs. These
findings indicate that human eggs infected at fertilization with HIV
will express viral proteins.
DE beta-Galactosidase/GENETICS Animal DNA, Viral/ADMINISTRATION &
DOSAGE/GENETICS Female *Gene Expression Genes, Reporter Human
HIV/*GENETICS HIV Infections/MICROBIOLOGY *HIV Long Terminal Repeat
In Vitro Lac Operon Mice Microinjections Ovum/MICROBIOLOGY Promoter
Regions (Genetics) Viral Proteins/GENETICS Zygote/MICROBIOLOGY
MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).