Effect of IPTG Concentration on Recombinant Human Prethrombin-2 Expression in Escherichia coli BL21(DE3) ArcticExpress

12th Congress of Indonesian Soc. for Biochemistry and Molecular Biology in Conjunction With The 2nd Int. Conf. "Collaboration Seminar of Chemistry and Industry

生物科学;化学

Silaban, S.^1 ; Gaffar, S.^2 ; Simorangkir, M.^1 ; Maksum, I.P.^2 ; Subroto, T.^2

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Negeri Medan, Medan

20221, Indonesia^1

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Sumenadg

45363, Indonesia^2

关键词: E. coli BL21(DE3);    expression;    Expression in Escherichia coli;    Gene transcriptions;    Inclusion bodies;    prethrombin-2;    Protein bands;    Soluble fraction;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/217/1/012039/pdf
DOI  :  10.1088/1755-1315/217/1/012039

学科分类：生物科学（综合）

来源: IOP

【 摘 要 】

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-β-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°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.