新型蛋白酶抑制剂基因hwtx-11在大肠杆菌中的克隆表达及杀虫功能研究  

Cloning,Expression of HWTX-11 Gene in E.coli and Its Bioactivity

作　　者：夏立秋[1] 曾智[1] 贺小红[1] 付祖姣[1] 单世平[1] 丁学知[1] 孙运军[1] 李文萍[1] 王海龙[1] 罗玉双[1] 

机构地区：[1]湖南师范大学生命科学学院微生物分子生物学湖南省重点实验室,中国长沙410081

出　　处：《湖南师范大学自然科学学报》2007年第4期90-93,共4页Journal of Natural Science of Hunan Normal University

基　　金：国家自然科学基金资助项目(30670052);国家"863计划"项目(2006AA02Z187;2006AA10A212);湖南省自然科学基金重点资助项目(06jj2009)

摘　　要：将化学合成的蜘蛛毒素蛋白酶抑制剂基因hwtx-11克隆至载体pGEX4T-1上,在大肠杆菌BL21(DE3)中进行表达.重组菌株在IPTG诱导下,经SDS-PAGE和Western blot分析表明,表达的GST-HWTX-11融合蛋白的相对分子量约为33 kD.对重组菌株诱导表达后的产物进行生物活性测定,GST-HWTX-11融合蛋白对甜菜夜蛾(Spodoptera exiguaHubner)和棉铃虫(Helicoverpa armigera)在3d时的LC50分别为86.6μg/mL、119.4μg/mL;其与Cry1Ac对甜菜夜蛾和棉铃虫幼虫毒力也有明显的协同作用.The gene of spider toxin, hwtx-11 has been synthesized by chemical method and cloned into the prokaryotic vector pGEX4T-1. The GST-HWTX-11 fusion proteins were expressed by IPTG induction and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） and Western blot with polyclonal antibody. The results showed that the molecular mass of the fusion protein was 33 kD. Bioassay showed that the LC50 against Spodoptera exigua Hubner and Helicoverpa armigera reached 86.6 μg/mL,119.4 μg/mL,respectively. More over, being synergism between GST-HWTX-11 and Cry1Ac against Spodoptera exigua Hubner and Helicoverpa armigera.

关 键 词：HWTX-11 融合表达 WESTERN BLOT 生物活性测定 

分 类 号：Q78[生物学—分子生物学]