大草蛉P450基因cDNA片段的克隆及吡虫啉诱导表达  

Cloning of Chrysopa septempunctata P450 cDNA Fragment and the Expression Induced by Imidacloprid

作　　者：刘昌燕[1] 曾凡荣[1] 

机构地区：[1]中国农业科学院植物保护研究所/农业部作物有害生物综合治理重点实验室,北京100081

出　　处：《中国生物防治学报》2014年第3期427-433,共7页Chinese Journal of Biological Control

基　　金：国家"948"计划项目(2011-G4);国家"973"计划项目(2013CB127600)

摘　　要：本研究根据不同昆虫细胞色素单加氧酶P450基因CYP4基因家族保守氨基酸序列,设计简并引物,通过RT-PCR扩增出了大草蛉Chrysopa septempunctata P450基因417 bp cDNA片段,命名为CSP450。该片段编码138个氨基酸残基,与已公布的烟草天蛾Manduca sexta、赤拟谷盗Tribolium castaneum、嗜卷书虱Liposcelis bostrychophila、玉米夜蛾Laphygma exigua、家蚕Bombyx mori、不吉按蚊Anopheles funestus和红火蚁Solenopsis invicta的CYP4 P450氨基酸序列进行比对,其相似性分别为59%、59%、54%、54%、58%、54%、54%。研究表明,用1 mg/L的吡虫啉溶液对大草蛉幼虫进行诱导,实时荧光定量PCR技术检测大草蛉体内CSP450基因的转录表达呈上调趋势,推测该基因可能参与了大草蛉体内吡虫啉的分解代谢过程。In this study, a pair of degenerate primers was designed based on the conserved amino acid regions of single oxygenase cytochrome P450 gene CYP4 gene family in insects. A P450 gene of 417 bp from Chrysopa septempunctata was amplified by reverse transcription polymerase chain reaction and named as CSP450. The cDNA fragment encoded 138 amino acid residues. The amino acid sequence was highly identical to those reported sequences of CYP4 P450 gene from Manduca sexca （59%）, Tribolium castaneum （59%）, Liposcelis bostrychophila （54%）, Laphygma exigua （54%）, Bombyx mori （58%）, Anopheles funestus （54%）, Solenopsis invicta （54%）. C. septempunctata larvae were treated by 1 mg/L imidacloprid. The transcription of C. septempunctata CSP450 gene in larvae was detected by real-time fluorescence quantitative PCR after imidacloprid induction. The results indicated that CSP450 expression was up-regulated by imidacloprid induction, demonstrating that CSP450 gene might participate in the catabolic process of imidacloprid in C. septempunctata.

关 键 词：大草蛉 P450基因 克隆 吡虫啉 诱导表达 

分 类 号：S476.2[农业科学—农业昆虫与害虫防治]