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dc.creatorHutapea, Hotma; Biomedical Research Center and Development Papua
dc.creatorOktavian, Antonius; Biomedical Research Center and Development Papua
dc.descriptionHuman 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.Key words: HIV-1,Iintegrase, E.coli JM109 AbstrakHIV (Human Immunodeficiency Virus) adalah virus RNA, dan termasuk ke dalam lentivirus, anggota kelompok retrovirus yang menyebabkan penyakit AIDS Acquired Immunodeficiency Syndrome. Tahap awal multiplikasi setiap retrovirus adalah integrasi genom DNA untai ganda virus ke dalam kromosom inang. Tahap integrasi ini difasilitasi oleh aktivasi enzim integrase. Tujuan penelitian ini adalah untuk memperoleh DNA pengkode integrase HIV. Kerangka baca terbuka atau Open Reading Frame (ORF) pengkode integrase yang diperoleh diklon ke vektor kloning pJETclone dan digunakan untuk mentransformasi Escherichia coli JM109. Sintesis cDNA dilakukan dengan mentranskripsi balik genom RNA menjadi DNA yang selanjutnya digunakan sebagai templat untuk amplifikasi ORF pengkode integrase dengan teknik reaksi polimerisasi berantai atau Polymerase Chain Reaction (PCR). Produk PCR selanjutnya disisipkan kedalam plasmid menggunakan sistem kloning blunt-ended dan dikarakterisasi dengan elektroforesis DNA dan analisis nukleotida. Analisis dengan elektroforesis DNA menunjukkan bahwa produk PCR berukuran sesuai dengan ukuran teoritis. Karakterisasi lebih lanjut dengan teknik analisis nukleotida menunjukkan produk PCR tersebut homolog dengan integrase HIV-1 dari Indonesia. Teknik PCR juga dilakukan terhadap klon, dan adanya produk PCR berukuran sesuai dengan ukuran teoritis menunjukkan adanya DNA sisipan. Analisis nukleotida dilakukan pada plasmid rekombinan murni dan DNA terbaca dengan baik.Keywords: HIV-1,Integrase, E.coli JM109en-US
dc.publisherPuslitbang Biomedis dan Teknologi Dasar Kesehatanen-US
dc.rightsThe Authors submitting a manuscript do so on the understanding that if accepted for publication, copyright of the article shall be assigned to Jurnal Biotek Medisiana Indonesia (The Indonesian Journal of Biotechnology Medicine) and Badan Penelitian dan Pengembangan Kesehatan (National Institute of Health Research and Development) as publisher of the journal.Copyright encompasses exclusive rights to reproduce and deliver the article in all form and media, including reprints, photographs, microfilms and any other similar reproductions, as well as translations. The reproduction of any part of this journal, its storage in databases and its transmission by any form or media, such as electronic, electrostatic and mechanical copies, photocopies, recordings, magnetic media, etc. , will be allowed only with a written permission from Jurnal Biotek Medisiana Indonesia (The Indonesian and Journal of Biotechnology Medicine) and Badan Penelitian dan Pengembangan Kesehatan (National Institute of Health Research and Development).Jurnal Biotek Medisiana Indonesia (The Indonesian and Journal of Biotechnology Medicine) and Badan Penelitian dan Pengembangan Kesehatan (National Institute of Health Research and Development), the Editors and the Advisory International Editorial Board make every effort to ensure that no wrong or misleading data, opinions or statements be published in the journal.
dc.sourceJurnal Biotek Medisiana Indonesia; Vol 2, No 2 (2013); 59-65id-ID
dc.sourceJurnal Biotek Medisiana Indonesia; Vol 2, No 2 (2013); 59-65en-US
dc.subjectHIV-1, Integrase, E.coli JM109en-US
dc.titleDiterima: 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 JM109en-US
dc.typePeer-reviewed Articleen-US

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