解毒酶基因cDNA克隆和高效表达

昆虫抗药性的一个重要机制是其产生的解毒酶可以将大剂量的农药脱去毒性1] 。酯酶活性升高是库蚊对有机磷杀虫剂抗性的主要机制 ,与酯酶Ｂ1有关的抗性最高2 ,3] 。酯酶与有机磷杀虫剂有非常强的结合力 ,可以迅速与之形成强结合体4 ] 。酯酶的解毒作用具有很高的手性专一性 ,在有机磷化合物 ,特别是高毒的有机磷化合物的解毒作用中非常重要1] 。高效表达解毒酶基因 ,将昆虫解毒酶用于人畜解毒的目的研究还未见报道。本文报道在大肠杆菌中高效表达昆虫解毒酶 ,并将产物用于实验动物有机磷中毒的解毒研究 ,为昆虫抗性相关基因的开发利用提…

Cloning and High Level Expression of Mosquito Detoxifying Gene

The full-length cDNA of mosquito esterase B1 had been isolated, and subcloned into pBV220. The recombinant vector pBV220B1 was constructed and transformed into E. coli DH5 alpha. A 60 kD protein was induced by 42 degrees C and its expression was temperature-dependent. After 6 h induction, the target protein occupied 50% of the total protein. The expressed product existed in both inclusion body and soluble proteins in the cells. The amount of the soluble detoxifying enzyme increased along with the induction time. The data of detoxifying experiments indicated that the detoxifying enzyme in expression strain of E. coli can detoxified toxicity of organophosphate insecticides, it showed a clear detoxifying affect on hens poisoned by organophosphate insecticides.