Tan Xin-yu, Fan Zheng, Wang Hua-jin, Shi Lei, Yin Bin, Ni An-ping, Qin Chuan, Zou Ke, Shen Yan, Yuan Jian-gang, Qiang Bo-qin, Peng Xiao-zhong
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 10005, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Oct;25(5):504-7.
To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain.
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.
Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained.
The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.
克隆、表达并纯化严重急性呼吸综合征冠状病毒（SARS-CoV）PUMC2株的核衣壳蛋白。
根据已发表的SARS-CoV基因组序列，通过逆转录聚合酶链反应（RT-PCR）克隆SARS-CoV PUMC2株N蛋白的全长cDNA，并将该cDNA克隆至pET32a表达载体。重组N蛋白在大肠杆菌BL21（DE3）中表达，并用镍离子-亚氨基二乙酸（Ni(2+)-NTA）进行纯化。
获得了原核表达并纯化的SARS-CoV PUMC2株N蛋白。
通过基因工程方法获得的SARS-CoV重组N蛋白可用于SARS-CoV N蛋白的进一步功能研究。
