title: [Preparation and characterization of polyclonal antibodies against haplotype chicken natural killer cell receptor B-NK19 and B-NK21].
abstract: OBJECTIVE
  To express natural killer (NK) cell inhibitory receptor gene B-NK of B19 and B21 haplotype chicken, which show different susceptible to various avian diseases, using prokaryotic expression system, and to prepare B-NK19 and B-NK21-specific rabbit polyclonal antibodies.
  
  
  METHODS
  B-NK genes of B19 (named as B-NK19) and B21 (named as B-NK21) chicken were amplified by reverse transcription PCR from peripheral blood lymphocytes of one B19 or B21 chicken, respectively, and then inserted into an expression vector of pET-30a. The recombinant plasmids of pET-B-NK19 and pET-B-NK21 were transformed into E.coli (DE3) and induced to express, respectively. Both His-tagged foreign proteins of B-NK19 and B-NK21 were seperated by SDS-PAGE and immunized to New Zealand rabbits to prepare polyclonal antibodies. The MHC-specificities of the sera were analyzed by Western blotting.
  
  
  RESULTS
  The chicken NK cell inhibitory receptor gene B-NK from MHC haplotypes B19 and B21 were successfully amplified, cloned and expressed in E.coli after induction with 1.0 mmol/mL IPTG for 4 h at 37°C, respectively. Western blotting revealed that the prepared rabbit serum antibodies positively reacted to the expressed foreign proteins.
  
  
  CONCLUSIONS
  Chicken NK cell inhibitory receptor proteins B-NK of B19 and B21 haplotypes could immunologically cross-react and their polyclonal antibodies are partially specific to B complex.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Fengming
authors.initials: F
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Huiqiang
authors.initials: H
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Chuanjiang
authors.initials: C
authors.lastname: Lian
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Laboratory Animal and Comparative Medicine Unit, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150001, China.
authors.forename: Lingxia
authors.initials: L
authors.lastname: Han
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D002645:Chickens; D003429:Cross Reactions; D006239:Haplotypes; D011817:Rabbits; D055607:Receptors, Natural Killer Cell / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology
chemical_list: D000906:Antibodies; D055607:Receptors, Natural Killer Cell; D011994:Recombinant Proteins