Functional Analysis of ktrA Gene Encoding Potassium Uptake Protein from Deinococcus radiodurans

doi:10.13304/j.nykjdb.2013.464

›› 2014, Vol. 16 ›› Issue (1): 76-81.DOI: 10.13304/j.nykjdb.2013.464

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Functional Analysis of ktrA Gene Encoding Potassium Uptake Protein from Deinococcus radiodurans

(1.Life Science and Engineering College, Southwest University of Science and Technology, Sichuan Mianyang 621010|2.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

Online:2014-02-15 Published:2014-02-17

耐辐射异常球菌K+吸收蛋白ktrA基因功能研究

鲜先毅1,2,江世杰1,2,崔广艳1,2,刘盈盈2,代其林1,陈明2,王劲2*

(1.西南科技大学生命科学与工程学院, 四川绵阳 621010|2.中国农业科学院生物技术研究所, 北京 100081)

通讯作者: 王劲，研究员，博士，主要从事微生物分子遗传学研究。E\|mail: wangjin@caas.cn
作者简介:鲜先毅|硕士研究生|研究方向为特殊环境微生物功能基因资源利用。E\|mail: xian_liu1314@163.com。
基金资助:
国家973计划项目(2013CB733903);国家863计划项目(2012AA063503);国家转基因生物新品种培育重大专项(2014ZX0801201B);农业部公益性行业科研专项(201103007);西南科技大学博士研究基金(11ZX7104)资助。

Abstract

Abstract:

The Deinococcus radiodurans R1 potassium uptake protein encoded by ktrA gene(DR\|166), played an important role under salt and osmotic stress. The quantitative real\|time PCR assay showed that the expression of ktrA gene was up\|regulated remarkably under salt and D\|sorbitol in wild strain. Construction of D. radiodurans R1 mutants deleted ktrA gene (ΔktrA) by means of fusion PCR and homologous genetic technology. Survival rates of the ΔktrA obtained and wild strain were investigated under different concentrations of KCl and D\|Sorbitol. The results indicated that the ΔktrA were much more sensitive to KCl and D\|Sorbitol treatment comparing to that of wild type, which indicated KtrA protein played an important role when influenced by salt and osmotic stress.This research will be helpful to study the functions of ktrA gene.

Key words: D. radiodurans R1, ktrA, potassium uptake protein, fusion PCR, abiotic stress

摘要：

耐辐射异常球菌(D. radiodurans R1)ktrA基因(DR_1666)编码的K+吸收蛋白在盐胁迫和渗透胁迫中起着重要作用。采用QRT\|PCR分析表明在盐和D\|山梨糖醇胁迫条件下，野生型菌株中ktrA基因表达均显著上调。同时采用融合PCR方法和同源重组技术，构建ktrA基因缺失突变株(ΔktrA)，以野生型菌株DR做对照，分别用不同浓度的氯化钾和D\|山梨糖醇对突变株进行胁迫处理。结果显示，突变株ΔktrA对氯化钾和D\|山梨糖醇的敏感性明显高于野生型，表明ktrA基因在菌株耐受高盐和高渗透胁迫时起重要作用。该结果为深入研究D. radiodurans R1菌株中ktrA基因的功能提供了很好的基础。

关键词: 耐辐射异常球菌；ktrA基因；K+吸收蛋白；融合PCR；非生物胁迫

CLC Number:

XIAN Xian\|yi1,2, JIANG Shi\|jie1,2, CUI Guang\|yan1,2, LIU Ying\|ying2, DAI Qi\. Functional Analysis of ktrA Gene Encoding Potassium Uptake Protein from Deinococcus radiodurans[J]. , 2014, 16(1): 76-81.

鲜先毅1,2,江世杰1,2,崔广艳1,2,刘盈盈2,代其林1,陈明2,王劲2*. 耐辐射异常球菌K+吸收蛋白ktrA基因功能研究[J]. , 2014, 16(1): 76-81.

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Functional Analysis of ktrA Gene Encoding Potassium Uptake Protein from Deinococcus radiodurans

耐辐射异常球菌K+吸收蛋白ktrA基因功能研究

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