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Publisher
IOP PUBLISHING LTD
DOI: 10.1088/1755-1315/217/1/012039
Keywords
E. coli BL21(DE3)ArcticExpress; expression; IPTG concentration; prethrombin-2
Funding
- grant Penelitian Terapan Unggulan Perguruan Tinggi (PTUPT) Year 2018 from Direktorat Jenderal Penguatan Riset and Pengembangan, Ministry of Technology Research and Higher Education, Republic of Indonesia
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Production of recombinant proteins in E. coli still facesa bottle neck due to formation of inclusion bodies. The level of gene transcription can be regulated by using appropriate concentration of isopropyl-beta-D-thiogalactoside (IPTG) inducer. The purpose of this study is to determine the effect of IPTG concentration on human prethrombin-2 (hPT-2) expression. The hPT-2 expression was induced by various concentrations of IPTG (0.01 mM, 0.025 mM, 0.05 mM, 0.075 mM, 0.1 mM, 0.2 mM and 0.3 mM) at 12 degrees C for 18 hours hosted by E. coli BL21(DE3) ArcticExpress. The result show that the suitable IPTG concentration for induction of hPT-2 in E. coli BL21(DE3) ArcticExpress was 0.1 mM. It was indicated from the 62-kDa protein band obtained from the soluble fraction on SDS-PAGE. It concluded that the IPTG concentration affect the rate of rhPT-2 expression.
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