细粒棘球绦虫核糖体蛋白S9基因克隆鉴定及其在不同发育阶段表达分析

发布时间：2018-06-10 14:18

本文选题：细粒棘球绦虫 + 核糖体蛋白S基因　；参考：《中国病原生物学杂志》2014年02期

【摘要】：目的克隆鉴定细粒棘球绦虫原头蚴核糖体蛋白S9基因,分析其在细粒棘球绦虫不同发育阶段的表达情况。方法从已构建的cDNA文库挑选基因,克隆EgRPS9基因并对编码产物的理化性质、功能位点、蛋白质的结构和功能域进行预测和分析,通过RealTime-PCR检测RPS9基因在细粒棘球绦虫不同发育阶段的表达情况。结果通过各种生物数据库对EgRPS9基因进行预测并测序,显示该基因具有完整开放阅读框,大小为564bp,编码188个氨基酸,预测分子质量单位为22.0ku,等电点为10.32。SMART保守功能区域分析发现该基因第107~149个氨基酸为40S亚基S9结构域S4超家族。通过多序列比对发现该基因具有较高的保守性,进化树结果表明该基因与曼式血吸虫、日本血吸虫及华氏睾吸虫亲缘关系较近。Real time-PCR检测原头蚴和囊泡RPS9基因相对表达量分别为1.16和1.02,差异无统计学意义(P0.05)。结论成功克隆出细粒棘球绦虫40S亚基RPS9新基因,该基因在细粒棘球绦虫不同发育阶段均有表达。
[Abstract]:Objective to clone and identify the ribosomal protein S9 gene of Echinococcus granulosus and analyze its expression in different developmental stages of Echinococcus granulosus. Methods the EgRPS9 gene was cloned from the constructed cDNA library, and the physicochemical properties, functional sites, structure and functional domain of the protein were predicted and analyzed. The expression of RPS9 gene in different developmental stages of Echinococcus granulosus was detected by RealTime-PCR. Results the EgRPS9 gene was predicted and sequenced by various biological databases. The result showed that the gene had a complete open reading frame with a size of 564bpand encoding 188 amino acids. The predicted molecular weight unit was 22.0ku.The isoelectric point was 10.32.SMART conserved functional region analysis showed that the 107-149 amino acids of the gene were the S4 superfamily of the 40S subunit S9 domain. It was found that the gene was highly conserved by multiple sequence alignment, and the result of evolutionary tree showed that the gene was associated with Schistosoma mansoni. The relative expression levels of RPS9 gene were 1.16 and 1.02 respectively in Procariae and vesicles of Schistosoma japonicum and Clonorchis Fahrensis. There was no significant difference between them (P 0.05). Conclusion A novel gene of Echinococcus granulosus 40S subunit RPS9 was successfully cloned and expressed in different developmental stages of Echinococcus granulosus.
【作者单位】：新疆医科大学;新疆医科大学基础医学院;新疆医科大学第一附属医院;
【基金】：国家自然科学基金项目(No.81000746) 新疆医科大学第一附属医院科研专项基金资助项目(No.2010YX01)
【分类号】：R383.33

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