Diterima: 8 April 2013 Direvisi: 6 Mei 2013 Disetujui: 20 Agustus 2013 59 Kloning Fragmen DNA Pengkode Integrase (Int) HIV (Human Immunodeficiency Virus) 1 Pada Escherichia Coli JM109

Abstract

Human Immunodeficiency Virus (HIV) is an RNA virus. It is a lentivirus, a retrovirus member family which causes Acquired Immunodeficiency Syndrome (AIDS). An early event in every retroviral multiplication is the integration of the viral double-stranded DNA genome into the host chromosome. The integration is facilitated by the activation of integrase.The goal of this research is to obtain the HIV integrase encoding gene. The obtained integrase ORF was intended to be cloned into cloning vector pJETclone and to transform Escherichia coli JM109. The cDNA synthesis was conducted by reverse transcription by converting the RNA genome to DNA. The obtained cDNA was amplified by Polymerase Chain Reaction (PCR) technique to obtain integrase encoding gene. The PCR product was inserted into the plasmid using blunt ended cloning system and was characterized using DNA gel electrophoresis and nucleotide analysis. The DNA gel electrophoresis of PCR product showed the expected band. Further characterization was conducted using nucleotide sequencing showed that the PCR product was homologue to HIV-1 integrase from Indonesia. The PCR was performed on the cloned showed DNA insert on expected size. The nucleotide analysis was conducted on the pure recombinant plasmid, the DNA was read properly