肥城桃果实VDAC基因的克隆、生物信息学分析及蛋白诱导表达  

Clone,Bioinformatics Analysis and Protein Induced Expression of VDAC Gene in Feicheng Peach Fruits

作　　者：衣冠帅 刘雨辰[1] 朱树华[1] 

机构地区：[1]山东农业大学化学与材料科学学院,山东泰安271018

出　　处：《山东农业科学》2016年第4期1-7,共7页Shandong Agricultural Sciences

基　　金：国家自然科学基金项目(31270723)

摘　　要：线粒体膜通透性转换孔(MPTP)是线粒体膜上的一种蛋白复合体,在线粒体介导的程序性死亡过程中起着重要作用。电压依赖型阴离子通道蛋白(VDAC)是MPTP的重要组成蛋白,可调节其开放与关闭,但VDAC在植物体中的作用机理尚不明确。本试验通过RT-PCR结合DNA末端快速扩增(RACE)方法,克隆得到肥城桃果实线粒体VDAC基因cDNA,全长共1 225 bp,其中编码区828 bp,编码276个氨基酸;氨基酸序列比对和系统进化树分析发现,桃果实VDAC与其它植物的VDAC蛋白具有较高的一致性;体外构建了pET-30a-VDAC重组表达载体,并成功在大肠杆菌BL21(DE3)中进行表达;使用镍柱纯化法,得到纯化的重组蛋白,为下一步研究桃果实VDAC蛋白的结构和功能奠定了基础。Mitochondrial membrane permeability transition pore（ MPTP） is a kind of protein complex,and plays important roles in cell mitochondria- mediated apoptosis. As an important part of MPTP,the voltage- dependent anion channel protein（ VDAC） regulates the opening of MPTP. However,its regulation mechanism in plant is still unclear. In this study,a c DNA encoding VDAC protein was isolated from Feicheng peach fruits by RT- PCR and rapid amplification of c DNA ends（ RACE）. The full- length c DNA of VDAC was 1 225 bp and contained 828 bp of coding sequence encoding a polypeptide of 276 amino acids. Multiple sequence alignment and phylogenetic tree analysis showed that the deduced protein VDAC of peach shared higher identify with that from other plants. The recombinant protein p ET- 30a- VDAC was constructed in vitro,expressed in Escherichia coli BL21,and purified through Ni- chelating affinity chromatography. The results would lay a foundation for studying the structure and function of VDAC protein in peach fruits in the next step.

关 键 词：肥城桃 线粒体 电压依赖型阴离子通道蛋白(VDAC) 基因克隆 生物信息学分析 蛋白表达 

分 类 号：Q945[生物学—植物学]