PROKARYOTIC EXPRESSION AND ANTIBODY PREPARATION OF THE EXTRACELLULAR DOMAIN OF DUCK TOLL-LIKE RECEPTOR 3
To study the function of duck TLR3, prokaryotic expression plasmid of Beijing duck Toll-like receptor 3 (Toll-like receptor 3, TLR3) gene was constructed and expressed in E.coli BL21 (DE3).The obtained TLR3 protein was purified from inclusion body and its polyclonal antibodies were prepared.The extracellular domain of duck TLR3 gene was amplified with PCR and the resulting 972 bp fragment was cloned into expression vector pET-32a (+).The recombinant plasmid pET-32a/TLR3 was then transformed into E.coli BL21(DE3) for expression with induction of IPTG.Duck TLR3 protein was purified from the obtained inclusion body.The recombinant protein was used to vaccinate rabbits to prepare polyclonal antibodies.The antiserum samples were collected and examined with Western blot and ELISA, demonstrating its specificity and reactivity.
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