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 DdrO in Deinococcus Radiodurans

作　　者：杜邱[1,2] 何淑雅[1,2] 马云[2] 李斌元[2] 孙晓宇[2] 廖端芳[3] 

机构地区：[1]南华大学公共卫生学院,湖南衡阳420001 [2]南华大学生物化学与分子生物学研究所,湖南衡阳420001 [3]南华大学药理教研室,湖南衡阳420001

出　　处：《现代生物医学进展》2011年第6期1037-1042,1071,共7页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30770647);湖南省自然科学基金(08JJ3033);湖南省科技厅科技计划项目(2010SK3036)

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。Objective：To clone the Deinococcus radiodurans ddrO gene,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 con-taining 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.

关 键 词：耐辐射球菌 ddrO基因 基因克隆 生物信息学 

分 类 号：Q937[生物学—微生物学] Q75