摘要
目的筛选结膜吸吮线虫基因组注释数据,鉴定海藻糖酶基因,并研究其编码蛋白的生物学特性。方法基于结膜吸吮线虫基因组筛选出海藻糖酶基因并进行PCR验证,通过生物信息学方法分析该基因编码蛋白的结构特征,包括疏水区、跨膜区、信号肽及结构域,以及编码蛋白质的二级、三级结构及其抗原表位。对其编码的氨基酸序列进行同源比对,利用MEGA X软件构建系统发育树,通过STRING数据库对其蛋白互作网络进行预测。结果从结膜吸吮线虫基因组中筛选鉴定出1条具有完整编码区的海藻糖酶基因序列,其长度为1638 bp,编码545个氨基酸;预测其编码蛋白大小为63478.48 ku,为分泌蛋白;其5′区域包含一个信号肽,无跨膜区,推测具有7个抗原表位。以STRING数据库中线虫蛋白互作网络信息为参考,对结膜吸吮线虫海藻糖酶基因的蛋白质互作网络进行预测,共鉴定出包括10个基因在内的27个互作关系。结论从结膜吸吮线虫基因组中筛选出1条海藻糖酶基因并对其编码蛋白进行了生物信息学分析,从而为该蛋白的生物学功能研究及结膜吸吮线虫病疫苗候选分子筛选奠定了基础。
Objective To characterize the trehalase gene in Thelazia callipaeda through screening the annotated data of the T.callipaeda genome,and to investigate the biological characteristics of the trehalase gene-coding protein.Methods The trehalase gene was screened from the T.callipaeda genome and subjected to validation by using a PCR assay.The structural features of the coding protein were analyzed with bioinformatics tools,including hydrophobicity,transmembrane region,signal peptides,conserved domains,as well as the secondary and tertiary structures and the antigen epitope.Homology analysis of the amino acid sequences was performed,and the phylogenetic tree was built by the MEGA X software.In addition,the protein-protein interaction network was deduced from the STRING database.Results The sequence of the trehalase gene with the complete CDS region was obtained from T.callipaeda genome,which had a length of 1638 bp and encoded 545 amino acids.The encoded protein was predicted to have a molecular weight of 63478.48 ku and be a secretory protein.The 5′domain of the encoded protein contained a signal peptide without transmembrane regions,and was predicted to contain 7 antigen epitopes.Based on the protein-protein interaction network of nematodes in the STRING database,the protein-protein interaction network of the trehalase gene of T.callipaeda was deduced,and 27 interactions covering 10 genes were identified.Conclusion A trehalase gene is successfully identified in T.callipaeda genome and its coding protein receives a bioinformatics analysis,which provides insights into the research on the biological functions of the protein and the screening of vaccine candidates for thelaziasis callipaeda.
作者
郑明辉
叶昌林
李先春
王莉君
沈玉娟
徐德林
ZHENG Ming-Hui;YE Chang-Lin;LI Xian-Chun;WANG Li-Jun;SHEN Yu-Juan;XU De-Lin(Zunyi Medical University,Zunyi 563003,China;Key Laboratory of Gene Diagnosis and Therapy of Education Office of Guizhou Province,China;Chinese Center for Disease Control and Prevention.China;Xinpu District Sub-bureau of Public Security,Zunyi Municipal Bureau of Public Security,Guizhou Province,China)
出处
《中国血吸虫病防治杂志》
CAS
CSCD
北大核心
2020年第1期60-68,共9页
Chinese Journal of Schistosomiasis Control
基金
国家自然科学基金(81760373)
中国博士后科学基金(2019M650799)
贵州省科学技术厅科学技术基金计划(黔科合基础[20161168).
