细粒棘球绦虫DNA损伤修复蛋白Rad50的生物信息学分析  

Preliminary validation and bioinformatics analysis of protein Rad50 in Echinococcus granulosus

作　　者：赵金龙 许少全 周润 夏衣旦木·吐尼亚孜 麦尔哈巴·麦麦提艾力 李婧 杜蕴 吕国栋 赵军[1,3] ZHAO Jinlong;XU Shaoquan;ZHOU Run;Xiayidanmu·Tuniyazi;Maierhaba·Maimaitiaili;LI Jing;DU Yun;LV Guodong;ZHAO Jun(Collegeof Pharmacy,Xinjiang Medical University,Urumqi 830054,China;Xinjiang Medical University Jointly Established the State Key Laboratory of Pathogenesis,Prevention,and Treatment of Diseases Highly Prevalent in Central Asia;The First Affiliated Hospital of Xinjiang Medical University,Department of Pharmacy,Xinjiang Key Laboratory of Clinical Drug Research)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830054 [2]新疆医科大学省部共建中亚高发病成因与防治国家重点实验室 [3]新疆医科大学第一附属医院药学部新疆药物临床研究重点实验室

出　　处：《中国病原生物学杂志》2024年第11期1305-1311,共7页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.82160700);新疆维吾尔自治区“天山英才”医药卫生高层次人才培养项目(No.TSYC202301A051)。

摘　　要：目的克隆细粒棘球绦虫DNA损伤修复蛋白Rad50(DNA damagerepair protein Rad50 of Echinococcus granulosus,EgRad50)基因全长序列,并进行生物信息学分析。方法利用PCR技术克隆EgRad50基因全长序列,并通过NCBI蛋白数据库获取EgRad50的氨基酸序列,利用ProtParam、ProtScale、SignalP 5.0、TMHMM和ProtComp 9.0预测EgRad50蛋白的理化性质、MotifScan和NetPhos 3.1预测翻译后修饰位点、DNAStar和IEDB预测抗原表位、CDD预测结构域、SOPMA预测二级结构、Swiss-Model和VMD构建三级结构、DNAMAN进行多序列分析、MEGA构建系统进化树。结果克隆获得EgRad50基因全长序列共4073 bp,生物信息学预测EgRad50蛋白具有亲水性,分子质量为156.45 ku,无信号肽裂解位点,无跨膜区域,定位于细胞核和细胞质。二级结构以α螺旋为主,约占77.41%。EgRad50蛋白包括ATP酶相关(ATPases associated,AAA)特征结构域、DNA双链断裂修复ATP酶结构域和ATP结合盒(ATP-binding cassette,ABC)结构域。多序列比对发现该蛋白与多房棘球绦虫同源性为92.70%,与其他物种同源性较低。系统发育树表明EgRad50与智人、小鼠等哺乳动物Rad50亲缘关系较远。结论成功克隆并鉴定EgRad50基因,EgRad50蛋白主要参与DNA修复、细胞进程以及端粒维持等生物过程,是DNA损伤修复途径的重要蛋白。Objective The bioinformatics analysis was conducted on the cloned full-length sequence of the DNA damagerepair protein Rad5o gene from Echinococcus granulosus.Methods PCR was used to clone the whole EgRad50 gene sequence.Subsequently,the amino acid sequence of EgRad50 was retrieved from the NCBI protein database.The physicochemical properties of EgRad5o were predicted using ProtParam,ProtScale,SignalP5.O,TMHMM,and ProtComp9.O.Additionally,phosphorylation sites were predicted by MotifScan and NetPhos3.1,antigenic epitopes were predicted using DNAStar and IEDB,and the domain was identified through the CDD database.The secondary structure was predicted by SOPMA,while the tertiary structure was constructed using Swiss-Model and Verify3D.Multiple sequence analysis was carried out with DNAMAN and VMD,and a phylogenetic tree was constructed using MEGA.Results The full-length 4073 bp of EgRad50 gene was cloned and uploaded to NCBI database(PP789594).Bioinformatics analysis predicts that the EgRad5o protein exhibits hydrophilic properties,with a molecular weight of 156.45 ku.It lacks a signal peptide cleavage site and transmembrane region,indicating its localization within the nucleus and cytoplasm.The protein's secondary structure is primarily composed ofα-helices,accounting for approximately 77.41%of the structure.Furthermore,it comprises the ATPases associated characteristic domain,DNA double-strand break repair ATP enzyme Rad50 domain,and ATP-binding cassette-Rad50 domain.Additionally,Multiple sequence alignment revealed that the homology of this protein with E.multilocularis was 92.70%,with lower homology with other species.The phylogenetic tree indicates that EgRad50 is closely related to Rad50 in mammals such as Homo sapiens and mice.Conclusion The EgRad50 gene was cloned and identified successfully.The results of bioinformatics analysis and prediction show that EgRad5o protein is mainly involved in biological processes such as DNA repair,cellular processes,and telomere maintenance.It is an important protein in th

关 键 词：细粒棘球绦虫 DNA损伤修复蛋白Rad50 生物信息学分析 

分 类 号：R383.33[医药卫生—医学寄生虫学]