【摘要】 从牛皮蝇Ⅱ期幼虫中提取总RNA,进行RT-PCR扩增牛皮蝇Hypodermin A基因。将扩增基因进行克隆测序,构建原核表达载体pET30-HA,转化大肠杆菌BL21(DE3)。用IPTG诱导表达,得到表达量达到全菌蛋白35%以上的特异性蛋白。SDS-PAGE分析表明,表达产物大部分以包涵体形式存在。Western-blot检测结果表明,获得的重组蛋白为重组Hypodermin A,具有较好的免疫活性。更多还原

【Abstract】 Total RNA was extracted from the second stage larva of H.lineatum.Hypodermin A gene of H.lineatum was amplified by RT-PCR.The target gene was cloned and sequenced.The prokaryotic expression vector pET30-HA was constructed and then transformed to E.coli BL21(DE3).The recombinant E.coli was induced and expressed by IPTG.The expression level of the specific protein was more than 35% of total protein.It was mainly present in inclusion body observed by SDS-PAGE.Western blot analysis indicated that the obtained protein was recombinant hypodermin A,and had good reactiogenicity against positive serum.更多还原