Expression and purification of SARS coronavirus membrane protein

Dai Wuxing; Lei Mingjun; Wu Shaoting; Chen Zhihao; Liang Liang; Pan Hujrong; Qin Li; Gao Shitong; Yuan Shishan; Zhang Renli

doi:10.1007/BF02831095

Current Medical Science ›› 2004, Vol. 24 ›› Issue (5) : 414 -416. DOI: 10.1007/BF02831095

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Expression and purification of SARS coronavirus membrane protein

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Abstract

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed intoEscherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-β-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.

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Dai Wuxing, Lei Mingjun, Wu Shaoting, Chen Zhihao, Liang Liang, Pan Hujrong, Qin Li, Gao Shitong, Yuan Shishan, Zhang Renli. Expression and purification of SARS coronavirus membrane protein. Current Medical Science, 2004, 24(5): 414-416 DOI:10.1007/BF02831095

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