[Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein].
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.
Tan
Xin-yu
XY
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 10005, China.
Fan
Zheng
Z
Wang
Hua-jin
HJ
Shi
Lei
L
Yin
Bin
B
Ni
An-ping
AP
Qin
Chuan
C
Zou
Ke
K
Shen
Yan
Y
Yuan
Jian-gang
JG
Qiang
Bo-qin
BQ
Peng
Xiao-zhong
XZ
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D000086462:Coronavirus Nucleocapsid Proteins; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
D000086462:Coronavirus Nucleocapsid Proteins; D018076:DNA, Complementary; D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins