家蚕细胞色素P450基因Bmcyp6A8的克隆及序列与表达分析  

Molecular Cloning, Sequence and Expression Analyses of Bmcyp6A8,a Cytochrome P450 Family Gene of Silkworm,Bombyx mori

作　　者：曾媛琴[1,2] 米智[1] 隆耀航[1] 杜文华[1] 朱勇[1] 

机构地区：[1]西南大学生物技术学院,重庆400716 [2]西南大学生命科学学院,重庆400716

出　　处：《蚕业科学》2010年第3期421-427,共7页ACTA SERICOLOGICA SINICA

基　　金：重庆市自然科学基金项目(No.2008AB1026)

摘　　要：昆虫细胞色素P450第6亚家族(CYP6)氧化酶在对异源有毒物质的代谢中具有重要作用。采用生物信息学方法获得与果蝇CYP6亚家族基因cyp6A8同源的家蚕cyp6A8基因序列,预测该基因的开放阅读框(ORF)为1572bp,编码523个氨基酸,推定的蛋白分子质量为61.52kD,等电点为8.17。以家蚕5龄第3天幼虫头部cDNA为模板,用设计的特异引物PCR扩增出一条约1500bp的条带,大小与家蚕cyp6A8基因的ORF预测值接近,命名为Bmcyp6A8基因(GenBank登录号:GQ241737)。同源性分析结果:Bmcyp6A8基因与野桑蚕cyp6AE8基因的相似性为93%;与棉铃虫cyp6AE12基因的相似性为57%;与人cyp3A43基因的相似性为48%。芯片数据分析显示Bmcyp6A8基因在家蚕5龄第3天幼虫的头部、表皮与中部丝腺前部高量表达,与家蚕CYP6亚家族的其它横向同源基因不同的是,只有该基因在中部丝腺前部表达,推测其具有功能特异性。Insect oxidases of the cytochrome P450 subfamily 6 （CYP6） play an important role in the metabolism of exogenous toxic substances. The sequence of a silkworm （ Bombyx mori） gene homologous to Drosophila cyp6A8 was obtained by bioinformatics. It was predicted to have an open reading frame （ORF） of 1 572 bp encoding 523 amino acids. The deduced protein has a molecular weight of 61.52 kD and an isoelectric point of 8. 17. The cDNA of the head of silkworm larvae at day 3 of the 5th instar was used as the template for PCR amplification with specific primers. A band of about 1 500 bp was obtained. Its length was close to the predicted ORF of silkworm cyp6A8 gene, and was thus named Bmcyp6A8 （GenBank accession number. GQ241737）. Sequence analysis showed that it is more closely related to cyp6AE8 of Bombyx mandarina, with sequence identity of 93%, while its sequence identities with cyp6AE12 of Helieoverpa armigera and cyp3A43 of Homo sapiens are 57% and 48% respectively. Microarray data analysis showed that Bmcyp6A8 gene in silkworm larvae at day 3 of the 5th instar had high expression levels in head, integument and anterior part of the middle silk gland. Compared to other paralogous genes of the CYP6 subfamily in silkworm, only Bmcyp6A8 gene is expressed in anterior part of the middle silk gland, suggesting a functional specificity of this gene.

关 键 词：家蚕 细胞色素P450第6亚家族 基因克隆 序列分析 表达特征 芯片数据分析 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]