title: [Construction, expression and immunogenicity of EV71 multiepitope-mGITRL eukaryotic plasmid].
abstract: OBJECTIVE
  To construct an enterovirus 71(EV71) multiepitope-mGITRL eukaryotic plasmid and study its immunogenicity in BALB/c mice.
  
  
  METHODS
  We first designed and synthesized VP1' epigene containing two B cells and two T cells epitopes of VP1, and amplified mGITRL gene by PCR. The VP1' epigene and mGITRL gene were then cloned into the expression vector pIRES to construct the recombination plasmid pIRES-VP1'-mGITRL. The recombination plasmid was transfected into COS7 cells by liposome-mediated method. The protein expressions of VP1' and mGITRL were detected by Western blotting. BALB/c mice were immunized with pIRES-VP1'-mGITRL plasmid, and its serum antibody titer was measured by ELISA.
  
  
  RESULTS
  The recombination plasmid pIRES-VP1'-mGITRL was successfully constructed as demonstrated by sequencing. Western blot analysis indicated that the VP1'-mGITRL fusion protein was expressed in COS7 cells and muscle cells. After BALB/c mice were immunized with this plasmid, we detected the high titer of anti-VP1 antibody in serum.
  
  
  CONCLUSIONS
  VP1'-mGITRL fusion protein can be highly expressed in COS7 cells and muscle cells by the construction and transfection of the recombination plasmid pIRES-VP1'-mGITRL, and it could elicit the dramatic immune response in mice.
authors.affiliation: Department of Laboratory Medicine, Affiliated People's Hospital of Jiangsu University, Zhenjiang, China. maopen365@163.com
authors.forename: Zhen-wei
authors.initials: ZW
authors.lastname: Mao
authors.forename: Hai-bing
authors.initials: HB
authors.lastname: Liu
authors.forename: Dong
authors.initials: D
authors.lastname: Zheng
authors.forename: Hui-yong
authors.initials: HY
authors.lastname: Peng
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.forename: Zhao-liang
authors.initials: ZL
authors.lastname: Su
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D029821:Enterovirus A, Human / Q000235:genetics* / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D048069:Tumor Necrosis Factors / Q000235:genetics* / Q000276:immunology*
chemical_list: D000939:Epitopes; D011993:Recombinant Fusion Proteins; C478966:Tnfsf18 protein, mouse; D048069:Tumor Necrosis Factors