中国生物防治学报
中國生物防治學報
중국생물방치학보
CHINESE JOURNAL OF BIOLOGICAL CONTROL
2014年
3期
427-433
,共7页
大草蛉%P450基因%克隆%吡虫啉%诱导表达
大草蛉%P450基因%剋隆%吡蟲啉%誘導錶達
대초령%P450기인%극륭%필충람%유도표체
Chrysopa septempunctata%P450 gene%clone%imidacloprid%inducible expression
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。
本研究根據不同昆蟲細胞色素單加氧酶P450基因CYP4基因傢族保守氨基痠序列，設計簡併引物，通過RT-PCR擴增齣瞭大草蛉Chrysopa septempunctata P450基因417 bp cDNA片段，命名為CSP450。該片段編碼138箇氨基痠殘基，與已公佈的煙草天蛾Manduca sexta、赤擬穀盜Tribolium castaneum、嗜捲書虱Liposcelis bostrychophila、玉米夜蛾Laphygma exigua、傢蠶Bombyx mori、不吉按蚊Anopheles funestus和紅火蟻Solenopsis invicta的CYP4 P450氨基痠序列進行比對，其相似性分彆為59%、59%、54%、54%、58%、54%、54%。研究錶明，用1 mg/L的吡蟲啉溶液對大草蛉幼蟲進行誘導，實時熒光定量PCR技術檢測大草蛉體內CSP450基因的轉錄錶達呈上調趨勢，推測該基因可能參與瞭大草蛉體內吡蟲啉的分解代謝過程。
본연구근거불동곤충세포색소단가양매P450기인CYP4기인가족보수안기산서렬，설계간병인물，통과RT-PCR확증출료대초령Chrysopa septempunctata P450기인417 bp cDNA편단，명명위CSP450。해편단편마138개안기산잔기，여이공포적연초천아Manduca sexta、적의곡도Tribolium castaneum、기권서슬Liposcelis bostrychophila、옥미야아Laphygma exigua、가잠Bombyx mori、불길안문Anopheles funestus화홍화의Solenopsis invicta적CYP4 P450안기산서렬진행비대，기상사성분별위59%、59%、54%、54%、58%、54%、54%。연구표명，용1 mg/L적필충람용액대대초령유충진행유도，실시형광정량PCR기술검측대초령체내CSP450기인적전록표체정상조추세，추측해기인가능삼여료대초령체내필충람적분해대사과정。
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.
