title: [Over-expression in Escherichia coli and purification of nucleocaspid and membrane protein of SARS coronavirus].
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.
authors.affiliation: Beijing Institute of Biotechnology, Beijing 100850, China.
authors.forename: Yan-Ping
authors.initials: YP
authors.lastname: Yi
authors.forename: Chu-Fang
authors.initials: CF
authors.lastname: Li
authors.forename: Yu-Ling
authors.initials: YL
authors.lastname: Shi
authors.forename: Lin-Hai
authors.initials: LH
authors.lastname: Li
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Sheng-Qi
authors.initials: SQ
authors.lastname: Wang
authors.forename: Qing-Jun
authors.initials: QJ
authors.lastname: Ma
authors.forename: Cheng
authors.initials: C
authors.lastname: Cao
mesh_terms: 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*
chemical_list: D019590:Nucleocapsid Proteins; D015678:Viral Structural Proteins