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家蚕细胞色素P450基因CYP6AE2的分子克隆与序列分析
引用本文:艾均文,董元凌,孔卫青,杨金宏,刘劲,张永亮,张学松,柳照应,朱勇.家蚕细胞色素P450基因CYP6AE2的分子克隆与序列分析[J].蚕业科学,2007,33(3):380-386.
作者姓名:艾均文  董元凌  孔卫青  杨金宏  刘劲  张永亮  张学松  柳照应  朱勇
作者单位:1. 农业部蚕桑学重点开放实验室,重庆,400716;湖南省蚕桑科学研究所,长沙,410127
2. 农业部蚕桑学重点开放实验室,重庆,400716
基金项目:教育部博士学科点专项基金(编号20060635008),重庆市教委基金项目(编号040208)
摘    要:为了在分子水平上研究家蚕(Bom byx mori)细胞色素P450基因的特性,更好地探讨该基因与家蚕抗性的相关机制,依据已有的家蚕基因组数据,通过BLASTP比对,选择与抗性相关的6家族中含有预测基因数量最多的CYP6AE亚家族的1条序列,并设计1对引物,用RT-PCR方法克隆了家蚕细胞色素P450家族基因CYP6AE2(Gen-Bank登陆号:EF415296)。该基因的ORF为1 572 bp,编码523个氨基酸,推定的蛋白质分子质量为60.2 kD,等电点为8.50。将该基因的cDNA和家蚕基因组序列比对,结果表明该基因只有1个内含子;与已知的同亚家族基因CYP6AE1(欧洲防风草结网毛虫Depressaria pastinacella)、CYP6AE12(棉铃虫Helicoverpa armigera)编码的氨基酸序列比对,同源性同为53%。

关 键 词:家蚕  细胞色素P450基因  基因克隆  序列分析
文章编号:0257-4799(2007)03-0380-07
修稿时间:2007-03-28

Molecular Cloning and Sequence Analysis of a Novel Cytochrome P450 Gene, CYP6AE2, From Silkworm, Bombyx mori
AI Jun-Wen,DONG Yuan-Ling,KONG Wei-Qing,YANG Jin-Hong,LIU Jin,ZHANG Yong-Liang,ZHANG Xue-Song,LIU Zhao-Ying,ZHU Yong.Molecular Cloning and Sequence Analysis of a Novel Cytochrome P450 Gene, CYP6AE2, From Silkworm, Bombyx mori[J].Acta Sericologica Sinica,2007,33(3):380-386.
Authors:AI Jun-Wen  DONG Yuan-Ling  KONG Wei-Qing  YANG Jin-Hong  LIU Jin  ZHANG Yong-Liang  ZHANG Xue-Song  LIU Zhao-Ying  ZHU Yong
Institution:1. The Key SericuItural Laboratory of Agricultural Ministry, Chongqing 400716, China; 2. The SericuItural Research Institute of Hunan Province, Changsha 410127, China
Abstract:To study the molecular characteristics of cytochrome P450 genes from Bombyx mori and further inquire into the relationship between cytochrome P450 and resistance,according to the bioinformation of the genomic sequences database of Bombyx mori,a sequence was selected by BLASTP,which is a member of the largest subfamily CYP6AE that was predicted from resistance-related CYP6 family.A pair of primers were designed and a new gene named CYP6AE2(GenBank accession number: EF415296) was successfully cloned from the midgut mRNA of Bombyx mori by RT-PCR.Its ORF(open reading frame) is 1 572 bp in length and encodes 523 amino acid residues,with a deduced molecular weight of 60.2 kD and isoelectric point of 8.50.Alignment of the cDNA with the silkworm genomic sequences revealed that there is only 1 intron.Amino acid sequence analysis showed those of CYP6AE2,CYP6AE1 from Depressaria pastinacella and CYP6AE12 from Helicoverpa armigera share 53% indentity.
Keywords:Bombyx mori  Cytochrome P450 gene  Gene cloning  Sequence analysis
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