(西南大学植物保护学院 昆虫学及害虫控制工程重庆市市级重点实验室,重庆 400716)
摘要:
从桔小实蝇Bactrocera dorsalis (Hendel)成虫体内提取总RNA,利用RT-PCR和cDNA末端快速扩增技术获得了一个新的细胞色素P450基因cDNA序列全长。该基因经细胞色素P450基因命名委员会命名为CYP4D46(GenBank登录号:GU292422),其cDNA全长为1717 bp,包含1530 bp的完整开放阅读框(ORF),编码510个氨基酸,理论分子量约为58.40 kD,等电点为8.82。系统发育分析表明该基因与昆虫第4家族P450基因具有较高的同源性。实时定量PCR分析发现,CYP4D46基因在脂肪体中的相对表达量较高,分别是马氏管和中肠内的756倍和60倍,说明CYP4D46可能与脂肪体的重要生理功能相关。
关键词:
桔小实蝇;P450;基因克隆;组织特异性表达
Molecular cloning and tissue specific expression profile of a novel P450 gene encoding CYP4D46 from Bactrocera dorsalis (Hendel)
HUANG Yong, SHEN Guang-Mao, LIU Li, XIONG Sai, JIANG Hong-Bo, WANG Jin-Jun* (Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Chongqing 400716, China)
Abstract:
In this study, the full length of a novel P450 gene from the oriental fruit fly Bactrocera dorsalis (Hendel) was cloned by using the RT-PCR and rapid amplification of cDNA ends (RACE) technology. This gene was named as CYP4D46 (GenBank Accession No: GU292422) by the P450 nomenclature committee. The full-length cDNA contains an open reading frame of 1530 bp encoding 510 amino acids. The putative protein of CYP4D46 shows predicted molecular weights of 58.40 kDa with a theoretical pI of 8.82. Phylogenetic analysis showed a closer relationship of CYP4D46 with CYP4 members of other insects. Quantitative real-time PCR showed that the transcripts of CYP4D46 were highest in the fat body, in which the relative quantity was 756 and 60 folds to the Malpighian tube and midgut, respectively. The tissue-specific expression implied that CYP4D46 was related to the physiological functions of fat body in B. dorsalis.
Key words:
oriental fruit fly; Cytochrome P450; gene cloning; tissue-specific expression
|