Expression and functional identification of recombinant SARS-CoV-2 receptor binding domain (RBD) from E. coli system
Xiangzheng Gao, Shanshan Peng, Shengsheng Mei, Keying Liang, Muhammad Saleem Iqbal Khan, Eu Gene Vong, Jinbiao Zhan
Journal：PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
IF：2.16
DOI：10.1080/10826068.2021.1941106
PMID：34187304
Published：2021-06-30
research field：线粒体生物学细胞生物学神经药理学神经退行性疾病天然产物研究
Abstract
The receptor binding domain (RBD) of SARS-CoV-2 is located in the C-terminal of S1 subunit of the spike (S) protein which is responsible for recognizing and binding to the angiotensin-converting enzyme 2 (ACE2) receptor. The DNA encoding the SARS-CoV-2 RBD was inserted into pET-28a (+) to construct expression plasmid pET-28a (+)/RBD. The desired RBD protein was produced in E. coli Rosetta (DE) and purified by a Ni-NTA column. The recombinant RBD was analyzed by SDS-PAGE and Western blot. The flow cytometry analysis indicated that the recombinant RBD is capable of binding to human ACE2 (hACE2) in the ACE2-overexpressed HEK293A-hACE2 cells. Our results demonstrated that recombinant RBD expressed in E. coli Rosetta (DE) strain has bioactivities and can be used as an antigen for diagnosis and as a tool for the development of novel anti-viral drugs against SASR-CoV-2.
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