大草蛉p450基因cdna片段的克隆及吡虫啉诱导表达.pdfVIP

30(3)427-433 中国生物防治学报 Chinese Journal of Biological Control 2014 年 6 月 大草蛉P450基因cDNA片段的克隆及吡虫啉诱导表达 刘昌燕，曾凡荣* （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 ） 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。 关 键 词：大草蛉；P450 基因；克隆；吡虫啉；诱导表达 中图分类号：S476.2 文献标识码：A 文章编号：1005-9261 (2014)03-0427-07 Cloning of Chrysopa septempunctata P450 cDNA Fragment and the Expression Induced by Imidacloprid * LIU Changyan, ZENG Fanrong (Key Laboratory of Integrated Pest Management in Crops, Ministry of Agriculture/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China) Abstract: 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%), S