2003•PubMedRequires access
[Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein].
Xin yu Tan, Zheng Fan, Hua jin Wang, Lei Shi, Bin Yin, An ping Ni, Chuan Qin, Ke Zou, Yan Shen, Jian gang Yuan, Bo-qin Qiang, Xiao zhong Peng
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Abstract
OBJECTIVE: To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain. METHODS: According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA. RESULTS: Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained. CONCLUSIONS: The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.