The Cloning Of Enolase Gene And The Analysis Of Its Expression In The Permethrin-resistance Associated C6/36Cell Strain By Using Rt-qpcr Technology

In this study, based on the EST fragment（GeneBank ID：EC093827.1）caused by thedifferent expression between resistant strains and susceptible strains of Culexquinquefasciatus, specific reverse transcription and amplification primers were designed, andby relying upon the approach of rapid amplification of cDNA ends (RACE), the full length ofthe enolase gene was amplifted from a resistant strain of Culex pipiens pallens. To analyzetheir bioinformatic characteristics, the enolase gene obtained in Culex pipiens pallens has1311bp, which is coding433amino acids, and it is a kind of hydrophilic protein with13phosphorylation sites. Homology alignment and Phylogenetic tree establishment of enolasebetween Culex pipiens pallens and other species indicated that enolase genes are highlyhomologous among diffrerent species. And the identity of enolase between Culex pipienspallens and Culex quinque fasciatus even reached to97.8%. It is also predicted that theconserved domains of enolase contained highly conserved animo acid sites. It is speculatedthat enolase gene has a certain correlation with the resistance of Culex pipiens pallens topermethrin and prelimimary forecasted the probably mechanisms of enolase gene in theresistance reaction. Then，by using the RT-qPCR technology, the expression difference ofenolase gene mRNA between permethrin-resistant C6/36cell strain andpermethrin-susceptible C6/36cell strain were detected and analysed. The result showed thatthe expression of enolase gene in permethrin-resistant C6/36cell strain was up-regulated,which was1.90times than in permethrin-susceptible C6/36cell strain.This study paves thefoundation for clarifying the resistance mechanism of the enolase gene and the invention ofnew pesticides.