抗辐射菌<italic style="font-style: italic">lexA</italic>基因的克隆及在大肠杆菌中表达<sup>*</sup>

王明锁; 杜泽吉; 孔向蓉; 苏燎原

您当前的位置：

首页 >

文章列表页 >

抗辐射菌lexA基因的克隆及在大肠杆菌中表达^*

研究论文 | 更新时间：2023-04-07

- 抗辐射菌lexA基因的克隆及在大肠杆菌中表达^*
- MOLECULAR CLONING AND EXPRESSION OF lexA GENE FROM THE RADIORESISTANT BACTERIUM Deinococcus radiodurans
- 辐射研究与辐射工艺学报 2000年18卷第3期页码：193-197
- 作者机构：
  
  1.苏州医学院放射医学系　苏州　215007
- 作者简介：
- 基金信息：
  
  卫生部优秀青年科技人才专项科研基金（1999年度批准）
- DOI：
  中图分类号：
扫描看全文
王明锁, 杜泽吉, 孔向蓉, 等. 抗辐射菌lexA基因的克隆及在大肠杆菌中表达^*[J]. 辐射研究与辐射工艺学报, 2000,18(3):193-197.

WANG Mingsuo, DU Zeji, KONG Xiangrong, et al. MOLECULAR CLONING AND EXPRESSION OF lexA GENE FROM THE RADIORESISTANT BACTERIUM Deinococcus radiodurans[J]. Journal of Radiation Research and Radiation Processing, 2000,18(3):193-197.
王明锁, 杜泽吉, 孔向蓉, 等. 抗辐射菌lexA基因的克隆及在大肠杆菌中表达^*[J]. 辐射研究与辐射工艺学报, 2000,18(3):193-197. DOI：

WANG Mingsuo, DU Zeji, KONG Xiangrong, et al. MOLECULAR CLONING AND EXPRESSION OF lexA GENE FROM THE RADIORESISTANT BACTERIUM Deinococcus radiodurans[J]. Journal of Radiation Research and Radiation Processing, 2000,18(3):193-197. DOI：

摘要

研究了旨在克隆抗辐射菌,Deinococcus radiodurans,的,lexA,基因并构建其表达载体，以便进一步研究,lexA,基因的功能及其在辐射抗性中的作用。主要的实验步骤包括分离提取抗辐射菌基因组DNA，分离出,lexA,基因并测定其序列，克隆于质粒载体PUC19, 用电击穿孔法将重组的质粒导入大肠杆菌JM109, SDS-PAGE检测基因的表达。用定点突变技术，改变,lexA,基因核糖体结合位点（RBS）, 以增强,lexA,基因的表达。结果表明，,lexA,基因位于基因组DNA的,BlnI-AscI,片断中，由630个碱基对（bp）组成，编码210个氨基酸（aa）, 理论上推定，分子量为22.5kD，等电点为6.4。用pUC19构建的,lexA,基因表达载体能在大肠杆菌JM109中表达，但表达效率低，改变,lexA,的核糖体结合位点RBS可提高该基因表达水平，使进一步分离纯化,lexA,蛋白质以及深入研究该基因的作用成为可能。

Abstract

In order to investigate the role of ,lexA, gene in the radioresistance of ,Deinococcus radiodurans, the expressing vector of ,lexA, gene was constructed. The genomic DNA was extracted from wild type ,Deinococcus radiodurans, KD8301. The ,lexA, gene was isolated from the genomic DNA and sequenced. The Plasmid vector pUC19 was used to clone ,lexA, gene and the electroporation was employed to transfer the recombinant plasmid into ,E. coli, JM109. SDS-PAGE was used to check the expression of ,lexA, gene. The ribosome binding site (RBS) of ,lexA, gene was mutated by means of Site -Directed Mutagenesis technique for the optimum of gene expression. The results indicated that the ,lexA, gene was located in the ,Blnl-AscI, fragment of ,Deinocococus radiodurans, genomic DNA, containing 630bp and coding 210 aa. The theoretically deduced molecular weight and the isoelectric point were 2.5KDa and 6.4 respectively. ,lexA, gene inserted into pUC19 could be expressed in JM109, but at very low level. After optimizing the RBS of ,lexA, gene, higher level of ,lexA, expression was obeserved. The results make it possible to isolate and purify ,lexA, protein, and further to investigate the function of ,lexA, gene in ,Deinococcus radiodurans.

关键词

抗辐射菌lexA基因基因克隆基因表达

Keywords

Deinococcus radioduransLexA geneGene cloningGene expression

references

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

耐辐射奇球菌DR_2327和DR_2328基因克隆及功能分析

p53通路相关基因表达作为辐射生物剂量计的研究现状

⁶⁰Co γ射线诱导人淋巴细胞线粒体Cyt b、ATPase6和ATPase8基因表达改变的研究

⁶⁰Co γ射线照射淋巴细胞诱导PIG3和GADD45基因mRNA变化

电离辐射对人外周血淋巴细胞GADD45基因表达的影响

抗辐射菌lexA基因的克隆及在大肠杆菌中表达*

MOLECULAR CLONING AND EXPRESSION OF lexA GENE FROM THE RADIORESISTANT BACTERIUM Deinococcus radiodurans

DOI：

摘要

Abstract

关键词

Keywords

references

抗辐射菌lexA基因的克隆及在大肠杆菌中表达^*