Plasmid Construction for Development of a High Throughput System Selection of New Anti-HIV Drugs Derived from Biological Resources Indonesia

摘要

AIDS (Acquired Immunodeficiency Virus) is a disease caused by the HIV virus (Human Immunodeficiency Virus) that attacks the body system. 99% of the causes of AIDS in the world caused by HIV-1. The most effective antiretroviral therapy method currently is HAART (Highly Active Antiretroviral Therapy). One of the therapy components in HAART is an HIV-1 protease inhibitor which has higher genetic barrier than the other HIV treatments. Various studies have been developed to find a novel anti-HIV drug that can be reached by most of the patients in low-limited settings, such as Indonesia. Therefore in this study we constructed a plasmid that can be used for developing a high throughput system as a selection of new anti-HIV drug candidates from Indonesian resources. For plasmid backbone, we used the P00201704939, a constructed plasmid that containing the AraC regulator gene and the GFP reporter gene. The HIV Id protease dimer domain was constructed in plasmid backbone and transformed into host cell Escherichia coli BL21 (DE3) using heat-shock method. Subsequently, transformation results were confirmed by using PCR and Sanger sequencing method. The PCR and sequencing results revealed that the HIV Id homodimer gene was successfully transformed into a backbone plasmid and there was no mutation in the nucleotide and amino acid sequences.