title: [Prokaryotic expression of extracellular region of human BTLA and preparation and identification of its antiserum].
abstract: OBJECTIVE
  To prokaryotically express and purify fusion protein containing extracellular region of human BTLA and prepare the antiserum of it.
  
  
  METHODS
  Human BTLA (hBTLA) gene was amplified by PCR, digested with enzymes, ligated and subcloned into a his-tagged prokaryotic expression vector to generate a recombinant plasmid named pET28a-hBTLA. Then pET28a-hBTLA was transformed into E.coli BL21 (DE3). The hBTLA fusion protein was obtained upon IPTG induction, purified by Ni-NTA Purification System, and analyzed by SDS-PAGE. Rabbit anti-hBTLA antiserum was prepared and identified.
  
  
  RESULTS
  The pET28a-hBTLA plasmid was confirmed to carry the correct hBTLA gene by sequencing. It expressed a 15.7kD protein that could be purified by Ni-NTA purification system. The titer of the multiclonal antibody was 1:16 detected by double diffusion test, and 1:20 000 by ELISA, respectively. The anti-hBTLA antibodies can bind to hBTLA specifically shown by Western blot.
  
  
  CONCLUSIONS
  The prokaryotic expression vector has been constructed successfully, leading to highly purified hBTLA protein. The antiserum of hBTLA has been prepared, with high titer and specificity.
authors.affiliation: Department of Clinical Immunology, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Shu-hua
authors.initials: SH
authors.lastname: Zheng
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Jing
authors.initials: J
authors.lastname: Lv
authors.forename: Hong-mei
authors.initials: HM
authors.lastname: Wang
authors.forename: Jing-jing
authors.initials: JJ
authors.lastname: Bao
authors.forename: Jun-fa
authors.initials: JF
authors.lastname: Xu
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D010957:Plasmids; D011817:Rabbits; D011971:Receptors, Immunologic / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: C482586:BTLA protein, human; D007106:Immune Sera; D011971:Receptors, Immunologic; D011994:Recombinant Proteins