title: [Construction of a prokaryotic expression vector of human tau multi-epitope peptide and immunogenicity of the expressed product].
abstract: OBJECTIVE
  To construct a prokaryotic expression vector of human tau multiepitope peptide for examining the immunogenicity of a TauP1/P2 DNA vaccine in mice using the expressed product.
  
  
  METHODS
  The coding sequence of Tau multiepitope peptide gene was amplified from the plasmid pVAX1-Tau by PCR and inserted into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant plasmid pGEX-4T-2-TauP1/P2. The positive recombinants were transformed into E.coli BL21 cells, and the expression of fusion protein GST-TauP1/P2 was induced by IPTG and identified by SDS-PAGE. Mice was immunized with TauP1/P2 DNA vaccine and the production of the specific antibodies was detected by Dot-blot analysis using the purified fusion protein.
  
  
  RESULTS
  A gene fragment 300 bp in length was amplified. Enzyme digestion and DNA sequencing verified correct construction of the prokaryotic expression plasmid pGEX-4T-2-TauP1/P2. The expression of target fusion protein GST-TauP1/P2 was detected by SDS-PAGE. Specific antibodies against TauP1/P2 were detected in the serum of mice immunized with the DNA vaccine using GST-TauP1/P2 fusion protein.
  
  
  CONCLUSIONS
  The constructed prokaryotic expression plasmid of human Tau multiepitope peptide is capable of expressing the target fusion protein, which specifically recognizes the specific antibodies against TauP1/P2 in mice immunized with TauP1/P2 DNA vaccine.
authors.affiliation: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, China. msunhaitao1988@yahoo.com.cn
authors.forename: Haitao
authors.initials: H
authors.lastname: Sun
authors.forename: Huaqiang
authors.initials: H
authors.lastname: Yang
authors.forename: Lujun
authors.initials: L
authors.lastname: Yang
authors.forename: Zhengyang
authors.initials: Z
authors.lastname: Li
authors.forename: Mouxuan
authors.initials: M
authors.lastname: Du
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Chen
authors.forename: Xiaodan
authors.initials: X
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D019444:Vaccines, DNA / Q000096:biosynthesis / Q000276:immunology; D016875:tau Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
chemical_list: D000939:Epitopes; D010455:Peptides; D011994:Recombinant Proteins; D019444:Vaccines, DNA; D016875:tau Proteins