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).