旋毛虫新生幼虫p46000抗原基因的表达与抗原性分析

Expression and Antigenicity Analysis of p46 000 Antigen fromNewborn Larvae of Trichinella spiralis

目的 对旋毛虫新生幼虫p460 0 0抗原基因进行原核表达并检测重组抗原的抗原性。 方法 利用聚合酶链反应 (PCR)扩增目的基因 ,克隆到原核表达载体 pET 2 8a ,构建重组表达质粒 ,经酶切及测序鉴定后转化大肠埃希菌BL2 1(DE3 ) ,以异丙基 β D 硫代半乳糖苷诱导表达。十二烷基磺酸钠 聚丙烯酰胺凝胶电泳 (SDS PAGE)、酶联免疫吸附测定 (ELISA)和蛋白质印迹法 (Westernblotting)分析表达产物。 结果 SDS PAGE结果显示表达产物的分子量约为Mr 480 0 0 ,与理论值相符。ELISA和Westernblotting结果表明 ,重组蛋白可被旋毛虫感染的猪血清和兔抗重组蛋白血清识别 ;兔抗重组蛋白血清可识别旋毛虫新生幼虫抗原Mr 460 0 0蛋白。 结论 成功表达了旋毛虫新生幼虫p460 0 0抗原基因 ,重组抗原具有良好的抗原性。

Objective To express and analyze p46 000 antigen from newborn larvae of Trichinella spiralis. Methods p46 000 antigen gene was subcloned to the pET28a expression system by PCR. The recombinant transformant was induced by IPTG and the antigenicity was analyzed with ELISA and Western blotting. Results The molecular weight of the expressed protein was about Mr 48 000 . ELISA and Western blotting showed that this recombinant protein could be recognized by T.spiralis infected swine serum and rabbit anti-recombinant protein serum, and the rabbit anti-recombinant protein serum could recognize a Mr 46 000 protein from newborn larvae of T.spiralis. Conclusion p46 000 recombinant antigen from newborn larvae of T.spiralis was expressed which shows a specific antigenicity.