title: [Expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and identification of its mimic antigenicity].
abstract: OBJECTIVE
  To study the expression of predicted B cell epitope peptide in S2 subunit of SARS coronavirus spike protein in E.coli and its mimic antigenicity to S2 protein.
  
  
  METHODS
  B cell epitopes in S2 subunit of SARS coronavirus spike protein was predicted using DNAStar software. The cDNA sequence encoding the B cell epitope peptide was constructed artificially by PCR and then cloned into the downstream of chaperone 10 gene in vector pET28a(+) to construct pET28-chap10-S2epi plasmid. The fusion protein, chap10-S2epi, was expressed in E.coli BL21(DE3) and identified by SDS-PAGE and Western blot. The rabbit was immunized by purified Chap10-S2epi for the preparation of antiserum, which was used to identify the mimic antigenicity of Chap10-S2epi to S2 protein by ELISA.
  
  
  RESULTS
  Chap10-S2epi fusion protein was successfully constructed and expressed in E.coli. The antiserum from the animal immunized by Chap10-S2epi recognized full length of SARS coronavirus S2 spike protein.
  
  
  CONCLUSIONS
  The predicted B cell epitope peptide of SARS coronavirus S2 spike protein can induce the antigenicity of S2 protein, which provides some fundamental data for developing engineering vaccine against SARS coronavirus infection.
authors.affiliation: Department of Molecular Biology, Basic Medical College, Jilin University, Changchun 130021, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Feng
authors.forename: Min
authors.initials: M
authors.lastname: Wan
authors.forename: Xuan-jun
authors.initials: XJ
authors.lastname: Wang
authors.forename: Pei-yin
authors.initials: PY
authors.lastname: Zhang
authors.forename: Yong-li
authors.initials: YL
authors.lastname: Yu
authors.forename: Li-ying
authors.initials: LY
authors.lastname: Wang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D018985:Epitopes, B-Lymphocyte; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV