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).