Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein.
- Author:
Xin-yu TAN
1
;
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
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Complementary; genetics; DNA, Viral; genetics; Escherichia coli; genetics; Genetic Vectors; Genome, Viral; Molecular Sequence Data; Nucleocapsid Proteins; biosynthesis; genetics; isolation & purification; RNA, Viral; genetics; Recombinant Fusion Proteins; biosynthesis; genetics; Reverse Transcriptase Polymerase Chain Reaction; SARS Virus; genetics; isolation & purification; Sequence Analysis, DNA
- From: Acta Academiae Medicinae Sinicae 2003;25(5):504-507
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain.
METHODSAccording 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.
RESULTSProkaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained.
CONCLUSIONSThe SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.
