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

  • Pavan Kumar Kalla
  • Lalnunthari K
  • Selvam Arjunan

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.

Keyword: GAG gene, HIV, RNA, Cloning, RT-PCR.

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How to Cite
Kalla, P. K., L. K, and S. Arjunan. “CLONING AND MOLECULAR CHARACTERIZATION OF GAG GENE FROM HIV-1 INTO E. COLI DH5A HOST”. International Journal of Current Pharmaceutical Research, Vol. 6, no. 4, 1, pp. 41-44, https://innovareacademics.in/journals/index.php/ijcpr/article/view/3394.
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