title: Prokaryotic expression and identification of B- and T-cell combined epitopes of Em95 antigen of Echinococcus multilocularis.
abstract: OBJECTIVE
  This study is to clone and identify B- and T-cell combined epitopes from Em95 antigen.
  
  
  METHODS
  The B- and T-cell combined epitopes were predicted using bioinformatic software. Two DNA sequences of Em95-1 (which contained the coding region of one B- and T-cell combined epitope) and Em95-2 (which contained the coding regions of two B- and T-cell combined epitopes) were amplified by PCR. Em95-1 and Em95-2 were cloned into pET32a vector for protein expression. Rabbit was immunized with the expressed proteins of rEm95-1 and rEm95-2 to produce polyclonal antibodies. The immunogenicity and antigenicity of rEm95-1 and rEm95-2 were examined by Western blot analysis.
  
  
  RESULTS
  The three B- and T-cell combined epitopes were successfully cloned and expressed in PET32a vector. The recombinant antigens of rEm95-1 and rEm95-2 could specifically bind the human serum from patients with alveolar echinococcosis and specifically bind the prepared polyclonal antibodies.
  
  
  CONCLUSIONS
  Three B- and T-cell combined epitopes were successfully cloned with good immunogenicity and antigenicity. Our data suggest that B- and T-cell combined epitopes predicted from the Em95 antigen may be used for the construction of high-valence vaccines and as targets for prevention of echinococcosis.
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Hongying
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Fengbo
authors.initials: F
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Xiumin
authors.initials: X
authors.lastname: Ma
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Haimei
authors.initials: H
authors.lastname: Ma
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Yuejie
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Xianfei
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Immunology, Xinjiang Medical University Urumqi 830011, Xinjiang, China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhu
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wen
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, China ; Department of Immunology, Xinjiang Medical University Urumqi 830011, Xinjiang, China.
authors.forename: Jianbing
authors.initials: J
authors.lastname: Ding
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000737:chemistry / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; C460870:EM95 protein, Echinococcus multilocularis; D000939:Epitopes; D011993:Recombinant Fusion Proteins
keywords: B- and T-cell combined epitopes; Em95 antigen; bioinformatics; prokaryotic expression