VALENSI
Jurnal Kimia VALENSI Volume 6, No. 1, May 2020

The Effect of Single Co-expression of The DnaK-DnaJ-GrpE and GroEL/ES Chaperones and Their Combination on Expression Intein-pretrombin-2 in Escherichia coli ER2566

Iman Permana Maksum (Biomolecular Health and Food Laboratory, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia, +62227794391)
D Agus Yusuf Wildan (Biomolecular Health and Food Laboratory, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia, +62227794391)
Khomaini Hasan (Medical School, Universitas Jenderal Achmad Yani, Jl. Ters. Jend. Sudirman, Cimahi, 022-6650646)
Toto Subroto (Biomolecular Health and Food Laboratory, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Indonesia, +62227794391)



Article Info

Publish Date
29 May 2020

Abstract

The use of recombinant thrombin in the manufacture of fibrin glue allows diseases contamination to be avoided. However, the expression of recombinant protein in E. coli still has a disadvantage of the formation of inclusion bodies, so it needs to be minimized by co-expression of chaperones. Therefore, the aim of this study was to determine the effect of single DnaK-DnaJ-GrpE and GroEL/ES chaperone expression and their combination on the expression of intein-pretrombin-2Ti,pH on E. coli ER2566. The method started with isolation of pTWIN1-prethrombin-2Ti,pH and pG-KJE8 from E. coli TOP10F' and DH5α respectively, the co-transformation of the expression host E. coli ER2566 using pG-KJE8 and pTWIN1-prethrombin-2Ti,pHvectors, the chaperone co-expression was induced using L-Arabinosa before IPTG induction and cell culture growth was incubated at 22 oC. The expression products were characterized by using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results of the co-expression of chaperone showed that the number of soluble fraction was higher than the one without co-expression of chaperone. In addition, the co-expression of chaperone using pG-KJE8 in intein-prothrombin-2Ti,pH expression was sufficient using tetracycline as an inducer.

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Journal Info

Abbrev

valensi

Publisher

Subject

Chemistry

Description

Jurnal Kimia Valensi is a biannual and peer-reviewed open access journal published by Department of Chemistry, Faculty of Science and Technology UIN Syarif Hidayatullah Jakarta. This journal covering all aspect of ...