[Over-expression in Escherichia coli and purification of nucleocaspid and membrane protein of SARS coronavirus].
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.
Yi
Yan-Ping
YP
Beijing Institute of Biotechnology, Beijing 100850, China.
Li
Chu-Fang
CF
Shi
Yu-Ling
YL
Li
Lin-Hai
LH
Li
Ping
P
Huang
Wei
W
Wang
Sheng-Qi
SQ
Ma
Qing-Jun
QJ
Cao
Cheng
C
D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D019590:Nucleocapsid Proteins; D015678:Viral Structural Proteins