title: [Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein].
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.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 10005, China.
authors.forename: Xin-yu
authors.initials: XY
authors.lastname: Tan
authors.forename: Zheng
authors.initials: Z
authors.lastname: Fan
authors.forename: Hua-jin
authors.initials: HJ
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.forename: Bin
authors.initials: B
authors.lastname: Yin
authors.forename: An-ping
authors.initials: AP
authors.lastname: Ni
authors.forename: Chuan
authors.initials: C
authors.lastname: Qin
authors.forename: Ke
authors.initials: K
authors.lastname: Zou
authors.forename: Yan
authors.initials: Y
authors.lastname: Shen
authors.forename: Jian-gang
authors.initials: JG
authors.lastname: Yuan
authors.forename: Bo-qin
authors.initials: BQ
authors.lastname: Qiang
authors.forename: Xiao-zhong
authors.initials: XZ
authors.lastname: Peng
mesh_terms: 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
chemical_list: D000086462:Coronavirus Nucleocapsid Proteins; D018076:DNA, Complementary; D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins