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棉铃虫细胞色素P450 CYP6B7基因的克隆与融合表达 被引量:6
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作者 马彩霞 李梅 +2 位作者 邱星辉 何凤琴 刘惠霞 《昆虫学报》 CAS CSCD 北大核心 2006年第6期944-949,共6页
细胞色素P450CYP6B7被推测与棉铃虫Helicoverpa armigera对拟除虫菊酯类杀虫剂的抗性有关,但至今尚无CYP6B7参与杀虫剂代谢方面的直接证据。为揭示CYP6B7的代谢功能,作者以棉铃虫幼虫基因组DNA为模板,以CYP6B7基因设计特异性引物,扩增... 细胞色素P450CYP6B7被推测与棉铃虫Helicoverpa armigera对拟除虫菊酯类杀虫剂的抗性有关,但至今尚无CYP6B7参与杀虫剂代谢方面的直接证据。为揭示CYP6B7的代谢功能,作者以棉铃虫幼虫基因组DNA为模板,以CYP6B7基因设计特异性引物,扩增出包含321bp内含子的CYP6B7基因。用反向PCR的方法消除内含子,获得包含完整的CYP6B7基因的开放阅读框。将CYP6B7基因与pMAL-c2X载体连接,并转化E.coliTB1细胞,在IPTG诱导下,CYP6B7能与载体基因编码的麦芽糖结合蛋白(MBP)在大肠杆菌中融合表达,表达产物经直链淀粉(amylose)柱亲和层析分离洗脱后,得到SDS-PAGE电泳纯的融合蛋白。 展开更多
关键词 棉铃虫 细胞色素P450 cyp687 基因克隆 融合表达
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抗氰戊菊酯棉铃虫细胞色素P450 CYP6B7基因的克隆及分析 被引量:3
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作者 唐涛 成玉红 +2 位作者 王成菊 张文吉 邱立红 《农药学学报》 CAS CSCD 2007年第4期370-375,共6页
以抗氰戊菊酯棉铃虫六龄幼虫中肠组织总RNA为模板,采用特异性引物,通过对反转录-聚合酶链式反应(RT-PCR)的条件进行不断探索和优化,成功克隆出全长为1 557bp的基因片段(GenBank登录号DQ497428)。该片段包括一个完整的开放阅读框架(1 515... 以抗氰戊菊酯棉铃虫六龄幼虫中肠组织总RNA为模板,采用特异性引物,通过对反转录-聚合酶链式反应(RT-PCR)的条件进行不断探索和优化,成功克隆出全长为1 557bp的基因片段(GenBank登录号DQ497428)。该片段包括一个完整的开放阅读框架(1 515bp)及5′端的42个碱基,编码504个氨基酸残基。与国外报道的细胞色素P450CYP6B7基因(GenBank登录号AF031468)的核苷酸、氨基酸同源性分别为97.75%和98.81%,为CYP6B7的等位基因。Northern杂交分析表明,抗性品系棉铃虫中肠组织中CYP6B7mRNA的表达量明显高于敏感品系的,初步表明CYP6B7在棉铃虫对氰戊菊酯的抗药性中起着重要作用。 展开更多
关键词 棉铃虫 氰戊菊酯 抗药性 细胞色素P450 cyp6b7 RT-PCR
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Identification of the 2-tridecanone cis-acting element in the promoter of cytochrome P450 CYP6B7 in Helicoverpa armigera 被引量:3
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作者 Li Xu Dong-Zhi Li +2 位作者 Yuan-Yuan Luo Jian-Ying Qin Li-Hong Qiu 《Insect Science》 SCIE CAS CSCD 2018年第6期959-968,共10页
The expression level of cytochrome P450 genes in insects can be induced by plant allelochemicals,which is important for insects to adapt to host plants.Cytochrome P450CYP6B 7has been reported to be involved in pyethro... The expression level of cytochrome P450 genes in insects can be induced by plant allelochemicals,which is important for insects to adapt to host plants.Cytochrome P450CYP6B 7has been reported to be involved in pyethroid insecticide resistance in Heli- coverpa armigera,and its transcription level was induced by some inducers.Currently,the regulatory mechanism of the induced expression of CYP6B7remains unknown,although it is very important for understanding the detoxification mechanism to allelochemicals in host plants.The objective of the present study was to investigate the eis-acting ele- ment in the promoter of CYP6B7 mediating the inducible up-regulation of CYP6B7in H.armigera by 2-tridecanone.The promoter region of CYP6B7was cloned by genome walking technique and analyzed by transient transfeetion assay.Progressive 5'deletion of the promoter region of CYP6B7revealed that the relative luciferase activity of construct -320/+232could be significantly induced by 2-trideeanone.Further stepwise deletion between -320 and -238 bp found that construct -292/+232 could also be significantly induced by 2-tridecanone,but the adjacent construct -256/+232could not,suggesting the essential role of the sequence between -292 and --257 bp for 2-tridecanone induction. Nucleotide mutations between -292 and -281 bp had no influence on the induction ef- fect by 2-tridecanone,but nucleotide mutations between -280 and -257 bp significantly decreased the induction effect.These results demonstrated that the cis-acting element for 2-trideeanone induction was between -280 and -257 bp in the promoter of CYP6B7. 展开更多
关键词 cis-acting element CYTOCHROME P450 cyp6b7 HELICOVERPA ARMIGERA induction 2-tridecanone
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