title: [Construction of a prokaryotic expression vector for apoptin and preparation of polyclonal antibody of apoptin].
abstract: OBJECTIVE
  To construct a prokaryotic expression vector for apoptin and prepare polyclonal antibody of apoptin.
  
  
  METHODS
  Apoptin gene amplified from pGEM-T/Apoptin plasmid by PCR was cloned into pET-28a (+). E.coli BL21 (DE3) was transformed by the recombinant plasmid, and apoptin protein expression induced by IPTG was analyzed by SDS-PAGE. BALB/c mice were immunized with the protein and the titer of the antibody was determined using indirect enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  Apoptin gene was successfully cloned into pET-28a (+), and the expression of a protein with relative molecular mass of about 17 000 was identified by SDA-PAGE. After 5 immunizations of the mice with the protein, the blood antibody titer reached 1:5x10(5).
  
  
  CONCLUSIONS
  The prokaryotic expression vector for apoptin is successfully constructed and the polyclonal antibody of apoptin is obtained, which allows further functional study of apoptin.
authors.affiliation: Department of Oncological Surgery, First Affiliated Hospital, Xi'an Jiaotong University College of Medicine, Xi'an 710061, China. wangjsh@mail.xjtu.edu.cn
authors.forename: Jian-Sheng
authors.initials: JS
authors.lastname: Wang
authors.forename: Ming-Xin
authors.initials: MX
authors.lastname: Zhang
authors.forename: Xiao-Yi
authors.initials: XY
authors.lastname: Duan
authors.forename: Fei
authors.initials: F
authors.lastname: Mo
authors.forename: Quan-Ying
authors.initials: QY
authors.lastname: Wang
authors.forename: Guang-Xiao
authors.initials: GX
authors.lastname: Yang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D016678:Genome; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids
chemical_list: D000906:Antibodies; D036022:Capsid Proteins; C091717:VP3 protein, Chicken anemia virus