title: [Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71].
abstract: OBJECTIVE
  To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.
  
  
  METHODS
  The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector. The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E. coli M15 with IPTG induction. After washing with 8 mol/L urea and purification with Ni-affinity chromatography, the recombinant proteins obtained were tested for immunogenicity by Western blotting and ELISA using rabbit antisera against enterovirus 71 and Coxsackie Virus A16.
  
  
  RESULTS
  The recombinant VP1-VP4 proteins were highly expressed in E. coli M15 and the purified proteins could be specifically recognized by the rabbit sera against enterovirus 71.
  
  
  CONCLUSIONS
  The expressed enterovirus 71 structural proteins show good immunogenicity and can be used for developing enterovirus 71 vaccine and detection kits.
authors: Pediatric Center, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors: Yuan-bin
authors: YB
authors: Song
authors: Nan
authors: N
authors: Yu
authors: Si-jie
authors: SJ
authors: He
authors: Xin-xin
authors: XX
authors: Chen
authors: Bin
authors: B
authors: Wang
authors: Xiao-yan
authors: XY
authors: Che
authors: Qi-yi
authors: QY
authors: Zeng
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D029821:Enterovirus A, Human / Q000235:genetics* / Q000276:immunology*; D004769:Enterovirus Infections / Q000821:virology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D055261:Immunogenetic Phenomena; D051379:Mice; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D036022:Capsid Proteins; D011994:Recombinant Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; C442459:VP2 protein, Foot-and-mouth disease virus; C442460:VP3 protein, Foot-and-mouth disease virus