›› 2012, Vol. 14 ›› Issue (5): 33-41.DOI: 10.3969/j.issn.1008-0864.2012.05.06

Previous Articles     Next Articles

Cloning and Bioinformatic Analysis of symrk Gene Cloned from Arachis hypogaea L.

DUAN Xiao-hong1,2, GENG Li-li2, SHU Chang-long2, ZHU Yan-ming1, ZHANG Jie2*   

  1. (1.College of Life Science, Northeast Agricultural University, Harbin 150030|2.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China)
  • Received:2012-03-26 Revised:2012-04-17 Online:2012-10-15 Published:2012-10-15

花生共生受体类蛋白激酶基因的克隆及生物信息学分析

段小红1,2,耿丽丽2,束长龙2,朱延明1,张杰2*   

  1. (1.东北农业大学生命科学学院, 哈尔滨 150030|2.中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193)
  • 通讯作者: 张杰,研究员,主要从事农业微生物与植物基因工程研究。Tel:010-62815921;E-mail:jzhang@ippcaas.cn
  • 作者简介:段小红|硕士研究生|研究方向为植物基因工程与分子生物学。E-mail:duanhong1013@163.com。
  • 基金资助:

    国家973计划项目(2009CB118902);国家863计划项目(2011AA10A203)资助。

Abstract:

Symbiosis receptor-like protein kinase (SYMRK) plays a key regulatory role in the symbiosis pathway of plant and microorganism. The full length and 560 bp upstream sequence of symrk gene were cloned from Arachis hypogaea Baisha1016 by genome-walking, respectively. Characters of the Ah-symrk and encoded amino acid residue sequence, including the general physical and chemical properties, transmembrane helices domains, localization sites in cells and senior structure, were predicted and analyzed by the methods of bioinformatics. The result showed that Ah-symrk contained 15 exons and 14 introns, and the cDNA full-length of Ah-symrk was 3 042 bp containing a complete ORF(2 781 bp), which encoded 926 amino acid. The protein kinase was a hydrophobic-acidic protein, which located in cell membrane. Three copies of root-motif-tapox1 related to root-specific expression were contained in upstream regulatory sequence. The result of RT-PCR showed that Ah-symrk was root-specific gene.

Key words: genome-walking, SYMRK, bioinformatics, RT-PCR

摘要:

共生受体类蛋白激酶(symbiosis receptor-like protein kinase, SYMRK)是控制植物与微生物共生的关键调控基因,利用基因组步移技术,克隆得到花生symrk的全长基因及其560 bp的ATG上游序列。采用生物信息学方法对该基因及其编码蛋白的常规理化性质、跨膜结构域、亚细胞定位和高级结构等进行了预测和分析,结果表明Ah-symrk由15个外显子和14个内含子组成,cDNA全长3 042 bp,包含2 781 bp的ORF,编码926个氨基酸,为疏水性酸性蛋白质,定位于细胞膜;ATG上游序列包含3个与根特异表达有关的顺式作用元件root motif tapox1;RT-PCR验证,该基因在花生根中特异性表达。

关键词: 基因组步移;共生受体类蛋白激酶;生物信息学;RT-PCR

CLC Number: