CLONING AND MOLECULAR CHARACTERIZATION OF GAG GENE FROM HIV-1 INTO E. COLI DH5A HOST

Abstract

Considering the worldwide increasing prevalence of human immunodeficiency virus type 1 (HIV-1) infection, World Health Organization (WHO) has intensified the access to the antiretroviral treatment. In spite of that one of the major issues to eradicate HIV-1 is the persistence of proviral human immunodeficiency virus type 1 (HIV-1) DNA reservoir. Although PCR detects HIV-1 DNA, the diagnosis of early, post exposure HIV infection prior to seroconversion can be achieved by the detection of proviral DNA by RTPCR. In the present study HIV-1 DNA were isolated from patient with HIV and using specific primer designed using Primer 3 plus software for the HIV-1 gag gene. The amplified gene was ligated with T vector and transformed into DH5αcells. The plasmid DNA obtained was then confirmed by restriction digestion and sequence analysis. The sequence was found to be 98% similar to that obtained in GenBank. Further research is required to express the gene to get the protein antigen for the production antibodies or effective vaccine for HIV-1. Considering the worldwide increasing prevalence of human immunodeficiency virus type 1 (HIV-1) infection, World Health Organization (WHO) has intensified the access to the antiretroviral treatment. In spite of that one of the major issues to eradicate HIV-1 is the persistence of proviral human immunodeficiency virus type 1 (HIV-1) DNA reservoir. Although PCR detects HIV-1 DNA, the diagnosis of early, post exposure HIV infection prior to seroconversion can be achieved by the detection of proviral DNA by RTPCR. In the present study HIV-1 DNA were isolated from patient with HIV and using specific primer designed using Primer 3 plus software for the HIV-1 gag gene. The amplified gene was ligated with T vector and transformed into DH5αcells. The plasmid DNA obtained was then confirmed by restriction digestion and sequence analysis. The sequence was found to be 98% similar to that obtained in GenBank. Further research is required to express the gene to get the protein antigen for the production antibodies or effective vaccine for HIV-1