[Expression of human Toll-like receptor-9 in E.coli and the preparation of antibody].
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.
Li
Jun
J
Department of Immunology, Southern Medical University, Guangzhou 510515, China. lijuns@fimmu.com
Zhao
Wen-zhong
WZ
Jiang
Hai-yan
HY
Zhu
Ping
P
Fu
Ning
N
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
D000906:Antibodies; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9