Research Article

Purification and Optimization of Prokaryotic Expression of CSN5B Protein of Arabidopsis thaliana  

Xu Niu1,2 , Lili Wang1,2 , Shenkui Liu3 , Yuanyuan Bu1,2
1 Key Laboratory of Saline-Alkali Vegetation Ecology Restoration (Northeast Forestry University), Ministry of Education, Harbin 150040, China
2 College of Life Science, Northeast Forestry University, Harbin 150040, China
3 The State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Lin’An, Zhejiang 311300, China
Author    Correspondence author
Molecular Soil Biology, 2020, Vol. 11, No. 2   
Received: 22 Jan., 2020    Accepted: 12 Feb., 2020    Published: 26 Feb., 2020
© 2020 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

CSN5B is a subunit of the COP9 signalosome (CSN) and plays physiological functions in the form of monomer or complex in plants. The CSN5B of Arabidopsis thaliana was cloned by RT-PCR technology and constructed into the prokaryotic expression vector pET-32a (+), which was introduced into E.coli BL21 host bacteria. Isopropylβ-D-1-thiogalactopyranoside (IPTG) was used to induce expression, and the cultural temperature and time was optimized. The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was employed to detect the induced recombination protein, and the protein purification was conducted. It was shown that the recombinant CSN5B introduced into E. coli could be induced to express, and the induction was relatively effective in the presence of 1mmol/L of IPTG at 30℃, 4h induction effect is better. The molecular weight of the induced recombinant protein was basically consistent with the theoretical molecular weight. This study provided a theoretical basis for in-depth further exploration of the functions of this gene in the future.

Keywords
CSN5B protein; Prokaryotic expression; Protein purification
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