Expression and purification of spike protein of severe acute respiratory syndrome coronavirus in Saccharomyces cerevisiae
- VernacularTitle:严重急性呼吸综合征冠状病毒刺突蛋白在酿酒酵母中的表达和纯化
- Author:
Lei YANG
;
Hong-Qin ZHANG
;
Shu-Zhen WU
;
Yun-Tian BI
;
Qi-Yu BAO
;
- Publication Type:Journal Article
- Keywords:
SARS virus;
Spike protein;
Reverse transcriptase polymerase chain reaction;
doning,molecular;
Gene expression;
Saccharomyces cerevisiae
- From:
Chinese Journal of Infectious Diseases
2007;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.
