Receptor-binding domain of SARS-Cov spike protein: soluble expression in E. coli, purification and functional characterization.

ABSTRACT:

Aim

Spike protein of coronavirus is responsible for virus binding, fusion and entry, and is a major inducer of neutralizing antibodies. This paper was to find a soluble and functional recombinant receptor-binding domain of severe acute respiratory syndrome-associated coronavirus (SARS-Cov), and to analyze its receptor binding ability.

Methods

Three fusion tags (glutathione S-transferase, GST; thioredoxin, Trx; maltose-binding protein, MBP), which preferably contributes to increasing solubility and to facilitating the proper folding of heteroprotein, were used to acquire the soluble and functional expression of RBD protein in Escherichia coli (BL21(DE3) and Rosetta-gamiB(DE3) strains). The receptor binding ability of the purified soluble RBD protein was then detected by ELISA

SUBMITTER: Chen J 

PROVIDER: S-EPMC4436633 | biostudies-literature | 2005 Oct

REPOSITORIES: biostudies-literature