论文

粟弯孢霉叶斑病菌G蛋白β亚基基因克隆及表达

  • 司贺龙 ,
  • 佟亚萌 ,
  • 郝志敏 ,
  • 李志勇 ,
  • 王楠 ,
  • 董志平 ,
  • 董金皋
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  • 1. 河北农业大学 生命科学学院, 河北 保定 071001;
    2. 河北省农林科学院 谷子研究所, 河北 石家庄 050035;
    3. 河北师范大学 生命科学学院, 河北 石家庄 050024
司贺龙(1975-),男,河北蔚县人,在读博士,主要从事分子植物病理学研究。

收稿日期: 2014-05-12

  网络出版日期: 2014-09-20

基金资助

谷子产业技术体系项目(CARS-07-12.5-A8);国家自然科学基金项目(31271787;31301616)

Molecular Cloning and Expression of G Protein β Subunit Gene of Curvularia lunata

  • SI He-long ,
  • TONG Ya-meng ,
  • HAO Zhi-min ,
  • LI Zhi-yong ,
  • WANG Nan ,
  • DONG Zhi-ping ,
  • DONG Jin-gao
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  • 1. College of Life Sciences, Agricultural University of Hebei, Baoding 071001, China;
    2. Millet Institute, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050035, China;
    3. College of Life Science, Hebei Normal University, Shijiazhuang 050024, China

Received date: 2014-05-12

  Online published: 2014-09-20

摘要

旨在获得粟弯孢霉叶斑病菌G蛋白β亚基基因,明确其表达模式,为阐明该基因对粟弯孢霉叶斑病菌致病性的调控机制奠定基础。运用SMART RACE RT-PCR技术,克隆粟弯孢霉叶斑病菌G蛋白β亚基基因 ClGβ 全长cDNA序列,以qRT-PCR 技术,对该基因在粟弯孢霉叶斑病菌不同生长时间的表达特征进行分析。结果表明,ClGβ 基因cDNA编码区为1 056 bp,DNA包含4个内含子和5个外显子,编码351个氨基酸。该基因的cDNA序列在GenBank中注册的登录号为JQ768316。qRT-PCR结果表明, 基因在菌株生长过程中表现出前期表达量较低而后期表达量明显升高的趋势。构建了pET28(a)-ClGβ 原核表达载体,经IPTG诱导,目的蛋白在宿主菌 E.coli BL21中获得表达,表达产物分子量与ClGβ蛋白计算分子量一致。

本文引用格式

司贺龙 , 佟亚萌 , 郝志敏 , 李志勇 , 王楠 , 董志平 , 董金皋 . 粟弯孢霉叶斑病菌G蛋白β亚基基因克隆及表达[J]. 华北农学报, 2014 , 29(4) : 7 -12 . DOI: 10.7668/hbnxb.2014.04.002

Abstract

In order to make a foundation for illustrating the pathogenic mechanism of G protein beta-subunit in Curvularia lunata,this research focuses on acquiring the gene encoding subunit in C.lunata and exploring its expression pattern.The full-length cDNA of G-protein beta-subunit was cloned using SMART RACE RT-PCR,and the expression pattern of ClGβ gene was analyzed under different growth time using Real-time PCR.The full-length cDNA of ClGβ was 1 056 bp and encoded 351 amino acids,while its DNA contained 4 introns and 5 extrons.The cDNA sequence of ClGβ had been deposited in GenBank with accession number JQ768316.The results of Real-time PCR indicated that the gene expression level of G-protein beta-subunit is lower at early stage and higher at later stage.We constructed the prokaryotic expression vector pET28(a)-ClGβ,and E.coli BL21 was transformed by this recombinant construct and induced by IPTG.The molecular weight of the expression product was identical with the calculated molecular weight of ClGβ.

参考文献

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