刚地弓形虫核糖体蛋白L35A基因克隆与生物信息学分析  

Cloning and bioinformatics analysis of ribosomal protein L35A of Toxoplasma gondii

作　　者：陈玉晴 卞安萍 张欣雨 许静雯 顾月月 李文超[1,2] 顾有方 刘欣超[1,2] CHEN Yuqing;BIAN Anping;ZHANG Xinyu;XU Jingwen;GU Yueyue;LI Wenchao;GU Youfang;LIU Xinchao(College of Animal Science,Anhui Science and Technology University,Fengyang 233100,China;Anhui Province Key Laboratory of Animal Nutrition Regulation and Health,Fengyang 233100,China)

机构地区：[1]安徽科技学院动物科学学院,安徽凤阳233100 [2]安徽省动物营养调控与健康重点实验室,安徽凤阳233100

出　　处：《安徽科技学院学报》2024年第6期9-16,共8页Journal of Anhui Science and Technology University

基　　金：安徽省高等学校自然科学研究项目(2022AH051641);安徽科技学院人才引进项目(DKYJ201902);安徽科技学院研究生科学研究项目(YK202109);大学生创新创业训练计划项目(202310879015,S202310879059,S202310879066)。

摘　　要：目的:克隆刚地弓形虫核糖体蛋白L35A基因,并运用生物信息学方法分析和预测RPL35A蛋白的结构与功能。方法:利用PCR技术扩增刚地弓形虫RPL 35 A基因,并通过ProtParam、ProtScale、TMpred和Signa IP预测RPL35A蛋白的理化性质、亲疏水性、跨膜区和信号肽等一级结构,通过MEGA 6.0和DANSTAR软件分析RPL35A蛋白的保守性。用NPSA SOPMA和Swiss-Model预测RPL35A蛋白的二级和三级结构。并通过PSORT、CD-Search、DUT和IEDB数据库预测RPL35A蛋白的亚细胞定位、结构域、翻译后修饰位点和抗原表位等。结果:刚地弓形虫RPL 35 A基因片段大小为339 bp,编码112个氨基酸,蛋白质分子式为C_(579)H_(940)N_(176)O_(151)S_(2),相对分子质量为12847.04,理论等电点为10.94,无跨膜区和信号肽,为亲水性不稳定蛋白,在不同虫株间高度保守。二级结构主要有无规则卷曲和延伸链。RPL35A蛋白亚细胞定位于线粒体,有9个磷酸化位点、12个O-糖基化位点和6个潜在的B细胞抗原表位。结论:刚地弓形虫RPL35A蛋白具有潜在的免疫原性,可作为刚地弓形虫疫苗和药物靶点研究的候选分子。Objective:To clone the ribosomal protein L35A of Toxoplasma gondii,and analyze the structure and function of RPL35A protein by bioinformatics.Methods:The RPL 35 A gene of T.gondii was amplified by PCR,and the primary structure of RPL35A protein,such as physical and chemical properties,hydrophobicity,transmembrane region and signal peptide,was predicted by ProtParam,ProtScale,TMpred and Signa IP.The conservation of RPL35A protein,was analyzed by using MEGA 6.0 and DANSTAR software.The secondary and tertiary structures of RPL35A protein were predicted by NPSA SOPMA and Swiss-Model.And the subcellular localization,structural domains,post-translational modification sites and antigenic epitopes of RPL35A protein were predicted by PSORT,CD-Search,DUT and IEDB databases.Results:The RPL 35 A gene of T.gondii was 339 bp,and encodes 112 amino acids,with a molecular formula C_(579)H_(940)N_(176)O_(151)S_(2).The relative molecular weight of RPL35A was 12847.04 and the oretical isoelectric point was 10.94.It had no transmembrane region and signal peptide,that was a hydrophilic unstable protein and highly conserved among different strains.The RPL35A protein was subcellularly localized in mitochondria with 9 phosphorylation sites,12 O-glycosylation s ites and 6 potential B-cell antigenic epitopes.Conclusion:The RPL35A protein of T.gondii was potentially immunogenic and could be used as a candidate molecule for T.gondii vaccine and drug target studies.

关 键 词：刚地弓形虫 核糖体蛋白L35A 结构 功能 生物信息学 

分 类 号：S855.9[农业科学—临床兽医学]