Stabilization and expression of HIVMN genes in Salmonella typhimurium.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Stabilization and expression of HIVMN genes in Salmonella typhimurium.

Abstr Gen Meet Am Soc Microbiol. 1994;94:151 (abstract no. E-46). Unique Identifier : AIDSLINE ASM94/94313067
Sizemore D; Branstrom A; Sadoff J; Warren R; Walter Reed Army Inst. of Res., Washington, DC.

Abstract: 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.

Keywords: 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 ABSTRACT
941030
M94A0887

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