桔小实蝇BdorGSTO1基因的克隆及发育表达分析

Cloning and Developmental Expression Pattern Analysis of GSTO1 from Bactrocera dorsalis Hendel.

为研究桔小实蝇(Bactrocera dorsalis Hendel.)Omega家族GST蛋白在发育过程的功能,采用RT-PCR和RACE技术克隆获得了桔小实蝇Omega家族GST基因的cDNA全长序列,命名为BdorGSTO1。该基因开放阅读框全长750bp,编码249个氨基酸,分子量约为28.52kD,等电点为6.46。氨基酸序列比对表明BdorGSTO1与刺舌蝇(Glossina morsitans morsitans)ADD18952的序列一致性最高,一致性值为70%,与豌豆蚜(Acyrthosiphonpisum)GST蛋白NP_001155757的一致性最低,一致性值为38.9%。荧光定量PCR分析表明BdorGSTO1在不同发育时期都有表达,且在1天蛹时表达量达到最高峰,随着蛹的发育其表达量逐渐降低,推测BdorGSTO1在桔小实蝇蛹的发育过程特别是在化蛹过程中发挥重要作用。

To explicit the function of Omega GST in the development process, a GSTgene belonging to Omega family was cloned using RT-PCR and RACE technology from Bactrocera dorsalis （Hendel） and was named as BdorGSTOI. The sequencing results showed that the open reading frame in BdorGSTO1 was 750 bp, encoding 249 amino acid residues. The protein molecular weight was about 28.52 kD and the isoelectric point of which was 6.46. Homology analysis of amino acid sequence showed that the BdorGSTO1 shared the highest sequence identity （70%） with ADD18952, and shared the lowest identity （38.9%） with NP_001155757 that from Acyrthosiphon pisum. Real-time PCR results showed that BdorGSTO1 expressed in whole developmental stages and the expression levels reached a peak in the 1 d pupae, which suggested that BdorGSTO1 played an important role in the development of pupae, especially in the pupation process.