title: [Amplification and cloning of the N gene of SARS-associated coronavirus].
abstract: OBJECTIVE
  To amplify and clone the N gene of severe acute respiratory syndrome-associated coronavirus.
  
  
  METHODS
  Using primer Premier 5.0 software, two pairs of nested PCR primers were designed to amplify the N gene. After purification, the amplified products were cloned into pMD18-T vectors, and the positive clones with the inserted fragments were identified by sequence analysis.
  
  
  RESULTS
  The amplified products was about 1 375 bp in length, and sequence analysis demonstrated that the N gene fragments had been successfully inserted into pMD18-T vectors.
  
  
  CONCLUSIONS
  The successful amplification and cloning of N gene facilitates further investigation of the expression of the N protein and study of its structure and functions.
authors.affiliation: Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
authors.forename: Wei-wei
authors.initials: WW
authors.lastname: Xiao
authors.forename: Wen-li
authors.initials: WL
authors.lastname: Ma
authors.forename: Bao
authors.initials: B
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Xiang-ming
authors.initials: XM
authors.lastname: Mao
authors.forename: Yi-fei
authors.initials: YF
authors.lastname: Peng
authors.forename: Yan-bin
authors.initials: YB
authors.lastname: Song
authors.forename: Qing-hua
authors.initials: QH
authors.lastname: Wu
authors.forename: Wen-ling
authors.initials: WL
authors.lastname: Zheng
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*
chemical_list: D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral; D019590:Nucleocapsid Proteins