生物学杂志

生物学杂志

• 研究报告 • 上一篇    下一篇

蛇床子RPS13蛋白的体外表达及生物信息学分析

  

  1. 1. 安徽中医药大学 新安医学教育部重点实验室 科研实验中心, 合肥 230038； 2．安徽中医药大学 药学院, 合肥230012； 3. 安徽道地中药材品质提升协同创新中心，合肥 230038； 4. 安徽省中医药科学院，合肥 230038
  • 出版日期:2018-04-18 发布日期:2018-04-18
  • 通讯作者: 吴家文，博士，教授，研究方向为中草药分子生物学，E-mail: wujiawen@ahtcm.edu.cn
  • 作者简介:王晨凯，硕士，研究方向为中草药分子生物学，E-mail: 846981810@qq.com
  • 基金资助:
    安徽省自然科学基金项目（1608085MH177）；安徽省留学人员科技活动启动项目（2015lx024）；安徽省高校优秀青年人才支持计划重点项目（gxyqZD2016139）；安徽中医药大学自然科学研究基金项目（2015zr017, 2015zr018）；安徽中医药大学人才引进科研项目（2015RC002）

Expression and bioinformatics analysis of RPS13 protein in Cnidium monnieri (L.) Cuss.

  1. 1. Key Laboratory of Xin′an Medicine, Ministry of Education, Anhui University of Traditional Chinese Medicine,Hefei 230038; 2. College of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230012;3. Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement, Hefei 230038; 4. Anhui Academy of Chinese Medicine, Hefei 230038, China
  • Online:2018-04-18 Published:2018-04-18

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摘要： 核糖体蛋白S13 ( Ribosomal protein S13, RPS13）属于核糖体蛋白家族,在核糖体中的生物学功能主要是维持核糖体大小亚基之间的动态连接。从蛇床子［Cnidium monnieri (L.) Cuss.］的根、茎、叶组织中提取总RNA，通过RT-PCR扩增蛇床子核糖体蛋白s13（rps13）的开放阅读框（ORF），成功获得了一段453 bp的基因，该基因片段编码151个氨基酸的蛋白；将获得的目的基因rps13 ORF连接到表达载体pET22b (+) 中，并进一步转化到大肠杆菌表达菌中诱导目的基因体外表达；通过对蛋白结构进行分析表明蛇床子RPS13含有6个 α 螺旋；对蛇床子RPS13进行系统进化分析表明该蛋白在进化上具有高度保守性，与植物类胡萝卜的RPS13亲缘关系最近。蛇床子RPS13蛋白在体外的成功表达以及对其进行系统的生物信息学分析为进一步深入研究该蛋白的结构和功能奠定了理论基础。

关键词: 蛇床子, rps13, 蛋白质表达, 生物信息学

Abstract: Ribosomal protein S13 (RPS13) is a key member of the ribosomal protein family, and plays an important role in the ribosome. In this study, total RNA was isolated from roots, stems and leaves of Cnidium monnieri (L.) Cuss. RT-PCR was used to obtain an open reading frame（ORF）of the rps13 which encoded 151 amino acid polypeptide; the target gene was cloned to pET22b(+) vector, then the recombinant plasmid was transformed into E. coli BL21 competent cells, and the fusion protein was expressed with induction of isopropyl β-D-1-thiogalactopyranoside (IPTG) and identified by Western Blotting. The three dimensional structure models showed that RPS13 contained six helices and the whole molecule appeared as a spiral structure with two clusters of helices connected by a long random coil, which may help to maintain the dynamic bridging interactions between the large and small subunits of ribosome. The phylogenetic analysis showed that the RPS13 of Cnidium monnieri (L.) Cuss. was highly conserved in evolution and most closely related to RPS13 of carrot. Expression and bioinformatics analysis of the RPS13 lay a foundation for the further study of its biological function.

Key words: Cnidium monnieri (L.) Cuss., ribosomal protein S13, protein expression, bioinformatics analysis