Cloning And Characterization Of Permethrin-resistant Associated Actin Gene And Cyp450Gene From Culex Pipiens Pallens

The Mosquito is a major media of virus diseases and parasitic diseases. Culex pipienspallens is a kind of the mosquito which spread many diseases in northern China.However,with the largely use of the pyrethroid pesticides, the mosquito produces differentlevels of resistance, greatly affected the insecticidal effect of the pyrethroid pesticides. Andthe mechanism of the development in resistance, is caused by the level of gene expressionchanges. Therefore, to handle the present situation of the drug resistance in mosquito, studyon the related genes which are related on the pesticide-resistence at molecular level is the keyof scientific and reasonable control of the mosquito.On the basis of the studies about the resistent-related genes on the mosquitos, thisresearch according to the obtained sensitivity express EST sequences from susceptible-strainand the resistant-strain by SSH technology combined with gene chip technology in Culexpipiens quinquefasciatus. Through the BLAST search, in GenBank database, searchg for therelated protein sequences expressed from the EST sequences. This study use the actin geneEST sequence and CYP450gene EST sequence in Culex pipiens quinquefasciatus asfoundation, designed specific gene primers, cloned the full-length resistant-related genes inCulex pipiens pallens,compared the genes' homology in these two species. Through RACEmethod, cloned the full length cDNA sequences, after bioinformatics analysis, preliminaryunderstand the biological characteristics, predict the resistance related mechanism, andexpressed the gene in E.Coli..This research preliminary obtained the full length cDNA sequences of gene actin andP450from the Culex pipiens pallens. The sequences analysis show that the whole length ofthe actin gene is1708bp, open reading frame for1131bp, coding the protein of376aminoacids. To get the gene bioinformatics analysis, the results show that the genetic code ofprotein has11hydrophobic areas and27phosphorylation sites; The total length of the P450gene is up to1791bp, open reading frame for1572bp, coding the protein of523amino acids.The series of bioinformatics analysis show that the genetic code of protein has14hydrophobic areas, protein conservative district is located in the amino acid sequence of the30-495place, and there are22phosphorylation sites. This study preliminary achieved the full length of the resistent-related genes, lay thefoundation on the further study of molecular mechanism,and to clarify the resistentmechanism and develop a new resistance related health pesticides.