用cDNA-RDA法克隆蚊虫抗药性基因

Insecticide-resistance is a major problem in controlling insect vectors of human diseases. Identification of genes associated with resistance or susceptibility to insecticides has been a valuable tool for understanding mechanisms underlying resistance to commonly used insecticides such as deltamethrin. .In this project（39970666）, 15 new resistance related genes （GenBank/NCBI BE247801、BE247803、BE247804、BE247806、BE247812、BE247823、BE247826、BE247828、BE247829、BE247832、BE247834、BE247837、BE247838、AY034058 and AY034059）were obtained in deltamethrin resistant versus susceptible laboratory strains of Culex pipiens pallens, with suppression subtractive hybridization (SSH), cDNA microarray and northern blots. Of these, 13 clones have sequence homology to known genes, such as ribosomal proteins, trypsin and chymotrypsin-like proteins. .In addition, Twenty-four new Cytochrome P450 genes (GenBank/NCBI AF157090-157093、AF160259、AF190775-190778 、AF190780-190789、AF191728-191730、AF208659和AF208660) were obtained from the two mosquito strains by RT-PCR,cDNA microarray and northern blots. They were approved by the Cytochrome P450 Nomenclature Committee, classified to the subfamilies CYP4C,CYP4D,CYP4H and CYP4J. Of these, 6 CYP4 genes may be related to deltamethrin resistance in Cx.pipiens pallens..The results may have important significance in the exploration of molecular mechanisms of pesticide resistance, the detection and management of resistance to pesticides, and the utilizations of pesticide-resistant genes, and mandate further investigation.

本研究拟采用cDNA-RDA法，从我们实验室选育的淡色库蚊对溴氰菊酯高纯抗性品系克隆抗性基因，并作结构分析。本研究的预期结果对于蚊虫拟除虫菊酯抗性的检测、抗性的分子机理以及抗性基因资源的利用等方面研究具有重要意义。