Cloning And Preliminary Studies On The Functions Of EU073017 Gene From Culex Pipiens Pallens

【Objective】To acquire EU073017 gene from Culex pipiens pallens and further research the relationship between EU073017 gene and insecticide resistance.【Methods】EU073017 gene was cloned from cDNA library of deltamethrin-resistant Cx.pipiens pallens,sequenced and analyzed with bioinformatic softwares.Transcriptions of EU073017 gene in the deltamethrin-susceptible strains and deltamethrin-resistant strains of Cx. pipiens pallens(fourth instar of larvae)was determined by Real-time PCR.The insect expression plasmids of EU073017 gene were constructed by gene reconstruction methods and transfected into Aedes albopictus C6/36 cells.Stable-transfected strains were established by screening culture with blasticidin.The viability of the transfected cells effected by various concentrations of deltamethrin were detected by Cell Counting Kit-8(CCK-8).【Results】EU073017gene were acquired with 398 bp including an CDS of 270 bp(GenBank no.EU073017,2007).Alignment of the deduced amino acid sequence with Anopheles funestus,An.gambiae and Ae. aegypti revealed sequence identities 86%,84%and 80%.EU073017 gene was overexpressed in deltamethrin-resistant strains than in deltamethrin-susceptible strains of Cx.pipiens pallens(fourth instar of larvae).Stable expression of EU073017 gene in C6/36 cells confer moderate deltamethrin resistantce.【Conclusion】EU073017 gene of Cx.pipiens pallens was cloned successfully and the gene is overexpressed at transcriptions level in the deltamethrin-resistant strains than in the deltamethrin-susceptible strains of Cx.pipiens pallens.EU073017 gene is related to deltamethrin resistance of the Culex mosquito.