title: [Expression of human Toll-like receptor-9 in E.coli and the preparation of antibody].
abstract: OBJECTIVE
  To express N terminal fragment of human Toll-like receptor-9 (hTLR-9) in E.coli and prepare its antiserum.
  
  
  METHODS
  The thioredoxin fusion expression plasmid of terminal fragment of hTLR-9 gene (707-1 167 bp) was constructed and then transformed into E.coli BL21 (DE3). The fusion protein was expressed in E.coli BL21 (DE3) under induction of 0.5 mmol/L IPTG. Inclusion body dissolved in urea was used as immunogen to prepare mouse anti-hTLR-9 antibody.
  
  
  RESULTS
  The fusion protein was expressed in the form of inclusion body with molecular mass of about 31kD. The purity of fusion protein reached to 90%; The result of immunohistochemical staining showed that the antisera against recombinant fusion protein could react specifically to HEK293 cells transfected with hTLR-9.
  
  
  CONCLUSIONS
  The fusion protein of hTLR-9 N-terminal fragment was expressed in E.coli BL21 (DE3). The prepared anti-hTLR-9 antibody can react specifically to HEK293 cells transfected with full length hTLR-9.
authors.affiliation: Department of Immunology, Southern Medical University, Guangzhou 510515, China. lijuns@fimmu.com
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Wen-zhong
authors.initials: WZ
authors.lastname: Zhao
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Jiang
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Ning
authors.initials: N
authors.lastname: Fu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D051217:Toll-Like Receptor 9 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D014162:Transfection
chemical_list: D000906:Antibodies; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9