Over-expression in Escherichia coli and purification of nucleocaspid and membrane protein of SARS coronavirus.
- Author:
Yan-Ping YI
1
;
Chu-Fang LI
;
Yu-Ling SHI
;
Lin-Hai LI
;
Ping LI
;
Wei HUANG
;
Sheng-Qi WANG
;
Qing-Jun MA
;
Cheng CAO
Author Information
1. Beijing Institute of Biotechnology, Beijing 100850, China.
- Publication Type:Journal Article
- MeSH:
Chromatography, Affinity;
Chromatography, Ion Exchange;
Enzyme-Linked Immunosorbent Assay;
Escherichia coli;
genetics;
metabolism;
Nucleocapsid Proteins;
genetics;
isolation & purification;
metabolism;
Reverse Transcriptase Polymerase Chain Reaction;
SARS Virus;
genetics;
metabolism;
Viral Structural Proteins;
genetics;
isolation & purification;
metabolism
- From:
Chinese Journal of Biotechnology
2003;19(4):392-396
- CountryChina
- Language:Chinese
-
Abstract:
Genes encoding nucleocaspid (N) and membrane (M) protein of SARS coronavirus were obtained by RT-PCR and were cloned into expression vector pET22b and pBV222. DNA sequencing showed that the genes cloned from a patient in Beijing were identical to the gene sequences from reported Toronto strain. The genes were over-expressed in E. coli either as inclusion body or as soluble form. The recombinant proteins were purified by ion-exchange, or ion-exchange followed by metal chelate affinity chromatography. The recombinant N protein was demonstrated highly antigenic and could be employed as antigen to detect SARS antibodies in ELISA system for SARS diagnosis.
