纹皮蝇Hypodermin B基因在大肠杆菌中的表达

Expression of Hypodermin B Gene of Hypoderma lineatum in Escherichia coli

从纹皮蝇期幼虫中提取总RNA,RT-PCR扩增纹皮蝇Hypodermin B(HB)基因.将扩增基因进行克隆测序,构建原核表达载体pET30-HB,转化大肠杆菌BL21(DE3).用IPTG诱导表达后,得到表达量达全菌蛋白30%以上的特异性蛋白.SDS-PAGE分析表明表达产物主要以包涵体形式存在.Western blotting检测结果表明,获得的重组蛋白为重组Hypodermin B,具有较好的反应原性.

Total RNA was extracted from the first instar Hypoderma lineatum larvae,then the Hypodermin B cDNA amplified by RT-PCR,was cloned and sequenced. The prokaryotic expression vector pET30-HB was constructed and transformed into E. coli BL21 （DE3）,then the recombinant E. coli was induced by IPTG. The expression level of the specific proteins was more than 30% of total proteins, and the expression product mainly presents in inclusion body observed by SDSPAGE. Western blotting analysis indicated that the obtained proteins were the recombinant of hypodermin B,and had good reactionogenicity against the positive serum.