摘要
线粒体膜通透性转换孔(MPTP)是线粒体内膜上的一种蛋白复合体,在细胞凋亡或坏死中具有重要作用。亲环蛋白D(Cy PD)是MPTP的重要组成蛋白,对线粒体引起的细胞凋亡有重要的调节作用,但植物中该蛋白的研究较少。本试验通过PCR、RACE等分子生物学技术克隆获得了肥城桃果实线粒体Cy PD基因c DNA序列,该序列全长663 bp,开放阅读框为615 bp,编码204个氨基酸;系统进化树分析发现,肥城桃Cy PD与苹果Cy PD蛋白在进化关系上最为亲密;蛋白质三维结构模拟表明,该蛋白含有2个α螺旋、8个β折叠结构;体外构建了p ET-32a-Cy PD重组表达载体,并成功在大肠杆菌BL21(DE3)中表达。这些结果为进一步研究Cy PD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 playing important roles in cell apoptosis and necrosis. Cyclophilin D（ CyPD） plays roles in cell mitochondria-mediated apoptosis,which is an important part of MPTP. However,there are fewer researches on CyPD in plants. In this study,the full-length c DNA of CyPD was gained by molecular biology method. It was 663 bp,contained 615 bp of the coding sequence and encoded a polypeptide of 204 amino acids. The recombinant CyPD protein was expressed in Escherichia coli. The phylogentic tree revealed that CyPD was more closely related to apple CyPD protein in evolution. The three-dimensional structure of CyPD showed that the protein contained 2 α-helix and 8 β-pleated sheets. These results would lay a foundation for studying the structure and function of CyPD protein further.
出处
《山东农业科学》
2017年第8期7-11,共5页
Shandong Agricultural Sciences
基金
国家自然科学基金项目(31270723)
