小地老虎P450CYP6AB基因的克隆与序列分析  

Molecular cloning and sequence analysis of CYP6AB from black cutworm, Agrotis ipsilon Hufngel

作　　者：樊东[1] 高艳玲[1] 邱广伟[1] 赵奎军[1] 周夏[1] 

机构地区：[1]东北农业大学农学院,哈尔滨150030

出　　处：《东北农业大学学报》2013年第7期137-142,共6页Journal of Northeast Agricultural University

基　　金：黑龙江省教育厅科学技术研究项目(12531045);黑龙江省研究生创新基金项目(YJSCX2011-071HLJ);国家现代农业产业技术体系建设专项基金项目(CARS-4)

摘　　要：细胞色素P450(Cytochrome P450或CYP450,P450)是一类广泛存在于细菌、真菌、植物和动物体内的亚铁血红素硫醇盐蛋白,该家族数量众多、功能复杂。在昆虫中,P450分布在昆虫的各个组织器官中,参与昆虫生长、发育、繁殖等重要生理活动,同时参与昆虫对植物毒素的适应性以及对常用化学杀虫剂的代谢。研究以小地老虎(Agrotis ipsilon Hufngel)5龄幼虫虫体为材料提取总RNA,利用RT-PCR和RACE技术,扩增得到小地老虎CYP6AB基因cDNA序列。该序列含有2 260个碱基,包括一个1 536个碱基的开放读码框,编码512个氨基酸,分子质量约为59.13 ku,多肽等电点为7.92。使用生物学软件分析发现,小地老虎CYP6AB蛋白序列中含有P450酶的特征结构FxxGxxxCxG(446-455)血红蛋白结合域。推导得到的氨基酸序列与其他昆虫,尤其是鳞翅目昆虫CYP6AB高度同源。获得的小地老虎CYP6AB基因cDNA序列已登录GenBank并获得登录号,登录号为JQ901386。Cytochromes P450 or CYP genes constitute one of the largest families of genes with representatives in virtually all living organisms, from bacteria to protists, plants, fungi and animals. The cytochreme P450 constitute a large and complex superfamily of heme-thiolate proteins. In insect, P450 enzymes are found in virtually all tissues, where they were involved in processes that were vital for insect growth, development and reproduction, as well as metabolism and detoxification of xenobiotics. P450 enzymes also played important roles in adaptation of insects to toxic compounds in their host plants and are involved in metabolism of all commonly used insecticides. CYP6AB cDNA was cloned by RT-PCR and rapid amplification of cDNA ends （RACE）. The cDNA was 2 260 base pairs in length, which contained an open reading frame of 1 536 base pairs coding for a polypeptide of 512 amino acid residues with a predicted molecular weight of 59.13 ku and pl 7.92. The putative protein contained the classic heme-binding sequence motif F ×× G ×× C × G （residues 446-455） which conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. The predicted protein shared extensive similarities with those from other insects, expecially the lepidopteran insects. The cDNA sequence had been deposited in GenBank with accession number JQ901386.

关 键 词：小地老虎 CYP6AB 克隆 序列分析 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治] Q785[农业科学—植物保护]