旋毛虫TS11-1丝氨酸蛋白酶抑制剂的克隆、表达及反应原性研究

he cDNA was tend to PCR using the primers designed according to the GENEBANK. The PCR production attached to 18T vector were sequenced and the most perfect one was reserved for the following assay.The target fragment was cut from 18T vector and connected to pET-28a expressing vector then turn to E.coli BL-21 and identified by PCR as well as dual-enzyme digestion.The IPTG was used to induce the expression on target protein.the results showed that the target protein expressed as inclusion body has the molecular weight of 42kDa.and the most perfect condition of induction was inducted along with 1mM IPTG at 37℃ for 5h.After emulsification with Freund's adjuvant,the recombinant protein was immunized to BALB/c mouse for four times to obtain polyclonal antibodies.then turn to Western-blot by antiserum antibody.the result showed that the Ts11-1 recombinant protein can be identified by polyclonal antibodies effectively and the stripe was single.All the results show that the TS11-1 recombinant protein has good immunogenicity and good reactogenicity.