Development of recombinant vaccine by cloning of gag, env and polrt genes of hiv ic in semliki forest virus vector

摘要

This invention relates to a process for the construction of a recombinant HIV-I vaccine comprising the steps of extraqting the proviral DNA, HIV1C from PBMC of individuals infected with HIV1-1, subjecting the individual genes gag env and polRT of HIVI C to PCR amplication, cloning the gag, env and polRT genes in the Semliki Forest virus vector, SFV2 gene expression vector and transformation in E. coli DHSo cells to obtain plasmids pSFV2 gen/gag, pSFV2 gen/env and pSFV2gen/polRT and pSFV-Helper2, growing said Plasmids in E. coli, purifying the plasmid DNA and linearizing the plasmid, DNA to,obtain the linearized SFV2gen recombinant plasmids, followed by in vitro transcription of the SFV2gen recombinant plasmids to obtain the RNAs, subjecting the RNAs to the step of co-transfection into BHK-21 cells along with SFV2-Helpet2 RNA to obtain recombinant SFV2gen particles, rSFV2gen/polRT.