EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI
Reverse transcriptase (RT) enzyme is a key tool in molecular biology for the synthesis of complementary DNA (cDNA) from messenger RNA (mRNA). Combining RT activity with PCR amplification has been a gold standard as the first step in cloning the coding region of any gene of interest. Evidently, RTs h...
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doaj-c05b46d50d1649849c43a1f745e99a422020-11-24T21:39:04ZengBogor Agricultural UniversityJurnal Ilmu Pertanian Indonesia0853-42172443-34622013-04-011814954EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLIUus Saepuloh0Diah Iskandriati1Joko Pamungkas2Dondin Sajuthi3Pusat Studi Satwa Primata, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Institut Pertanian Bogor, Jalan Lodaya II/5 Bogor 16151Pusat Studi Satwa Primata, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Institut Pertanian Bogor, Jalan Lodaya II/5 Bogor 16151Pusat Studi Satwa Primata, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Institut Pertanian Bogor, Jalan Lodaya II/5 Bogor 16151Pusat Studi Satwa Primata, Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Institut Pertanian Bogor, Jalan Lodaya II/5 Bogor 16151Reverse transcriptase (RT) enzyme is a key tool in molecular biology for the synthesis of complementary DNA (cDNA) from messenger RNA (mRNA). Combining RT activity with PCR amplification has been a gold standard as the first step in cloning the coding region of any gene of interest. Evidently, RTs have been critical in advancing molecular biology, genetics and medicine to their current stage. In this study, we were developing the RTRNase H recombinant enzyme isolated from serotype-2 simian betaretrovirus-2 (SRV-2) infected Indonesian Macaca fascicularis using Escherichia coli expression system. The study was conducted using RT SRV-2 gene expression using E. coli expression system, proteins purification, and application to RT PCR technique. The SDS PAGE expression analysis showed a specific band size of 32.7 kDa assumed as RT protein enzyme. Application of RT SRV-2 enzyme generated to the RT PCR technique of β-globin CDV and SRV-2 env gene target showed that the RT SRV-2 enzyme was capable to reverse transcribed mRNA into cDNA as indicated by the presence of specific DNA band compared with commercial RT enzymes. This RT SRV-2 enzyme showed its activity similar to that of commercial one, although the activity was lower. http://journal.ipb.ac.id/index.php/JIPI/article/view/8366escherichia coli expression systemrecombinant enzymereverse transcriptaseSRV-2 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Uus Saepuloh Diah Iskandriati Joko Pamungkas Dondin Sajuthi |
spellingShingle |
Uus Saepuloh Diah Iskandriati Joko Pamungkas Dondin Sajuthi EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI Jurnal Ilmu Pertanian Indonesia escherichia coli expression system recombinant enzyme reverse transcriptase SRV-2 |
author_facet |
Uus Saepuloh Diah Iskandriati Joko Pamungkas Dondin Sajuthi |
author_sort |
Uus Saepuloh |
title |
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI |
title_short |
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI |
title_full |
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI |
title_fullStr |
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI |
title_full_unstemmed |
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI |
title_sort |
ekspresi enzim rekombinan reverse transcriptase (rtrnase h) simian betaretrovirus serotipe-2 asal macaca fascicularis indonesia dalam sistem ekspresi eschericia coli |
publisher |
Bogor Agricultural University |
series |
Jurnal Ilmu Pertanian Indonesia |
issn |
0853-4217 2443-3462 |
publishDate |
2013-04-01 |
description |
Reverse transcriptase (RT) enzyme is a key tool in molecular biology for the synthesis of complementary DNA (cDNA) from messenger RNA (mRNA). Combining RT activity with PCR amplification has been a gold standard as the first step in cloning the coding region of any gene of interest. Evidently, RTs have been critical in advancing molecular biology, genetics and medicine to their current stage. In this study, we were developing the RTRNase H recombinant enzyme isolated from serotype-2 simian betaretrovirus-2 (SRV-2) infected Indonesian Macaca fascicularis using Escherichia coli expression system. The study was conducted using RT SRV-2 gene expression using E. coli expression system, proteins purification, and application to RT PCR technique. The SDS PAGE expression analysis showed a specific band size of 32.7 kDa assumed as RT protein enzyme. Application of RT SRV-2 enzyme generated to the RT PCR technique of β-globin CDV and SRV-2 env gene target showed that the RT SRV-2 enzyme was capable to reverse transcribed mRNA into cDNA as indicated by the presence of specific DNA band compared with commercial RT enzymes. This RT SRV-2 enzyme showed its activity similar to that of commercial one, although the activity was lower. |
topic |
escherichia coli expression system recombinant enzyme reverse transcriptase SRV-2 |
url |
http://journal.ipb.ac.id/index.php/JIPI/article/view/8366 |
work_keys_str_mv |
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