文章摘要
杜邱1,2 何淑雅1,2△ 马云2 李斌元2 孙晓宇2 廖端芳3.耐辐射球菌转录因子DdrO 的基因克隆与生物信息学分析[J].,2011,11(6):1037-1042
耐辐射球菌转录因子DdrO 的基因克隆与生物信息学分析
Cloning and Bioinformatics Analysis of Transcription Factor DdrOin Deinococcus Radiodurans
  
DOI:
中文关键词: 耐辐射球菌  ddrO 基因  基因克隆  生物信息学
英文关键词: Deinococcus radiodurans  ddrO  Gene cloning  Bioinformatics
基金项目:国家自然科学基金(30770647),湖南省自然科学基金(08JJ3033)和湖南省科技厅科技计划项目(2010SK3036)
作者单位
杜邱1,2 何淑雅1,2△ 马云2 李斌元2 孙晓宇2 廖端芳3 南华大学公共卫生学院 
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中文摘要:
      目的:克隆耐辐射球菌ddrO 基因,并对其进行生物信息学分析,预测其功能。方法:根据耐辐射球菌ddrO 基因序列,由 Primer Premier 5 设计一对引物,以提取的耐辐射球菌基因组为模板,PCR 扩增获得耐辐射球菌ddrO 基因,序列测定并利用生物 信息学软件对ddrO 基因的理化性质、高级结构及生物学功能等进行分析与预测。结果:成功获得了ddrO 基因。生物信息学分析 发现,ddrO 基因核苷酸序列长度为396bp,编码一个131aa 组成的相对分子质量为14.993kD 的预测的DdrO 转录因子。核酸同源 性搜索及比较分析仅在与耐辐射球菌同属的Deinococcus geothermalis 和Deinococcus deserti 中发现高度相似的序列;蛋白同源 性搜索发现一些与DdrO 显著同源的蛋白,如Deide_20570 (95%),Dgeo_0336 (90%),Deide_3p02170 (82%)等;结构域分析发现 DdrO 含有HTH (helix-turn-helix) DNA 结合结构域。结论:根据生物信息学结果预测DdrO 蛋白可能具有转录调控作用,参与 DNA 修复和复制,在耐辐射球菌的DNA 损伤修复过程中发挥一定作用。
英文摘要:
      Objective: To clone the Deinococcus radiodurans ddrOgene, and predict its function by bioinformatics analysis. Methods: According to the published ddrO gene sequence of Deinococcus radiodurans, by using the software of Primer Premier 5, a pair of primers were designed and synthesized. By using the genomic DNA isolated from Deinococcus radiodurans as templates for polymerase chain reaction (PCR), and Deinococcus radiodurans ddrO gene were gained. Sequenced and various bioinformatics softwares were employed to analyze and predict its physicochemical properties, advanced structure and biological function. Results: The ddrO gene was successfully obtained. Bioinformatics analysis revealed that ddrO nucleotide sequence length was 396bp and encoded a transcription factor DdrO containing 131 amino acid with a molecular weight of 14.993 KD. Nucleic acid homology search and comparative analysis showed that highly similar sequences were found only belong to Deinococcus geothermalis and Deinococcus deserti, which are the same genus with DR; some significant homology of the DdrO protein were found by Protein homology search, such as Deide_20570 (95%), Dgeo_0336 (90%), Deide_3p02170 (82%), etc.; and domain analysis showed that DdrO containing a HTH (helix-turn-helix) DNA-binding domain. Conclusion: Based on the results of bioinformatics, we predict that DdrO protein may have transcriptional regulatory function, possibly through a mechanism involved in the DNA repair and replication in Deinococcus radiodurans and played an important role in the process of DNA damage repair.
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