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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References


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About this article

Title

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

Date

07-10-2014

Additional Links

Manuscript Submission

Journal

International Journal of Current Pharmaceutical Research
Vol 6, Issue 4 (Oct-Dec 2014) Page: 41-44

Online ISSN

0975-7066

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Authors & Affiliations

Pavan Kumar Kalla

Lalnunthari K

Selvam Arjunan


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