title: [Expression of chicken pulmonary surfactant protein A and preparation of its polyclonal antibody].
abstract: OBJECTIVE
  To express chicken pulmonary surfactant protein A (cSP-A) in Escherichia coli and prepare polyclonal antibodies against the recombinant cSP-A.
  
  
  METHODS
  The carbohydrate recognition domain (CRD) gene of cSP-A (cSP-A-CRD) was artificially synthesized, subcloned into the prokaryotic expression vector pGEX-6P-1, and expressed in Escherichia coli BL21(DE3). After the expression products were identified by SDS-PAGE, the gel containing the target protein was cut and further purified. Mice were innoculated with the purified recombinant protein to prepare polyclonal antibodies against cSP-A. The antibody titer was determined by ELISA, and the specificity and application scope of the polyclonal antibodies were analyzed by Western blotting, immunohistochemical fluorescence assay and ELISA.
  
  
  RESULTS
  The serum against cSP-A was obtained and had an ELISA titer of 1×10(5);. The serum also showed excellent specificity and reactivity as demonstrated by the Western blotting. It could be used to detect the cSP-A in chicken lung epithelial cells by immunohistochemical fluorescence staining and could also be used to detect the hydrosoluble cSP-A in chicken bronchoalveolar lavage fluid by indirect ELISA.
  
  
  CONCLUSIONS
  The recombinant cSP-A was successfully expressed, and its mouse polyclonal antibodies were obtained.
authors.affiliation: School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Shao
authors.forename: Boyan
authors.initials: B
authors.lastname: Huang
authors.forename: Xiuhong
authors.initials: X
authors.lastname: Zhou
authors.forename: Ling
authors.initials: L
authors.lastname: Pan
authors.forename: Guijun
authors.initials: G
authors.lastname: Wang
authors.forename: Kezong
authors.initials: K
authors.lastname: Qi
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D002645:Chickens / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D008297:Male; D051379:Mice; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary; D037662:Pulmonary Surfactant-Associated Protein A / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
chemical_list: D000906:Antibodies; D037662:Pulmonary Surfactant-Associated Protein A