用mRNA RT-PCR差异显示技术克隆拟除虫菊酯抗性家蝇细胞色素P450基因片段

细胞色素P450单加氧酶系是一类广泛分布在生物有机体中的重要酶系,具有多样的功能。细胞色素P450介导的杀虫剂代谢解毒作用的增强是昆虫对杀虫剂产生抗性的普遍机制。为揭示家蝇对拟除虫菊酯类杀虫剂的分子基础,我们利用mRNA差异显示技术(DDRTPCR)从溴氰菊酯抗性品系家蝇Muscadomestica中分离出2个新的基因片段(M1,M2)。序列比对(BlastP)结果表明,M1与黑腹果蝇Drosophilamelanogaster中的Cyp6A9在氨基酸水平上具有54%的相似性,M2分别与果蝇Drosophilasimulans中的Cyp6g1有65%的相似性。M1和M2包含P450的特征序列,并与细胞色素P450具有最高程度的序列相似性,是为细胞色素P450基因片段。

CLONING OF CYTOCHROME P450 ESTs FROM A DELTAMETHRIN-RESISTANT STRAIN OF HOUSEFLY, MUSCA DOMESTICA, USING mRNA DIFFERENTIAL DISPLAY RT-PCR TECHNIQUE

Cytochrome P450-dependent monooxygenases are a very important Multiple-functional enzymatic system.The roles of cytochrome P450s in insect resistance to insecticides have been a main concern over the last decades.To explore the molecular basis of resistance of housefly to pyrethroid insecticides,we isolated two new cytochrome P450 fragments by using mRNA differential display RT-PCR technique from a laboratory housefly strain resistant to deltamethrin,designated M1(AY960133)and M2(AY960134).The deduced amino acid sequence of M1 shows 54% identity with Cyp6a24 of Drosophila melanogaster,and that of M2 shows 65% identity with Cyp6g1 from the Drosophila simulans.