家蚕细胞色素P450基因CYP6AE2的分子克隆与序列分析  

Molecular Cloning and Sequence Analysis of a Novel Cytochrome P450 Gene,CYP6AE2,From Silkworm,Bombyx mori

作　　者：艾均文[1] 董元凌[1] 孔卫青[1] 杨金宏[1] 刘劲[1] 张永亮[1] 张学松[1] 柳照应[1] 朱勇[1] 

机构地区：[1]农业部蚕桑学重点开放实验室

出　　处：《蚕业科学》2007年第3期380-386,共7页ACTA SERICOLOGICA SINICA

基　　金：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%。To study the molecular characteristics of cytochrome P450 genes from Bombyx moiland 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 subfamiry 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.

关 键 词：家蚕 细胞色素P450基因 基因克隆 序列分析 

分 类 号：Q963[生物学—昆虫学] Q78