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

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.