摘要
【目的】建立饱血状态雌性微小扇头蜱的microRNA文库,鉴定所含microRNA的种类,分析miR-275靶蛋白Vg-2的二、三级结构,检测其信号肽、跨膜结构域、磷酸化及糖基化位点,预测其优势抗原表位。【方法】以雌性、饱血状态的微小扇头蜱为研究对象,通过高通量测序技术建立其microRNA文库,利用软件miRDP及检索miRBase数据库,鉴定饱血状态雌性微小扇头蜱所含的microRNA种类;运用在线软件EXPASY、PRABI与SWISS-MODEL等分析Vg-2蛋白质理化性质,推断其二、三级结构;运用在线软件SignalP 5.0、TMHMM、NetPhos 3.1及NETCGlyc 1.0检测该蛋白的信号肽、跨膜结构域、磷酸化与糖基化位点;利用在线软件ABCpred Prediction、Scratch、IEDB和NetCTL预测Vg-2蛋白的B、T细胞优势抗原表位。【结果】获得成熟的microRNA 383个,其中67个为已被鉴定和注释的microRNA,316个为新发现的microRNA;微小扇头蜱的Vg-2基因序列全长5040 bp,共编码1679个氨基酸;Vg-2蛋白为亲水性酸蛋白,分子大小为189.89 kDa,理论等电点(pI)为6.08,其二级结构以无规则卷曲为主要结构成分,而其三级结构却以β-折叠为主要结构成分;Vg-2蛋白存在268个磷酸位点,20个糖基化位点,无信号肽和跨膜结构域,拥有58个B细胞优势抗原表位和8个T细胞优势抗原表位。【结论】在饱血状态雌性微小扇头蜱体内含有383个microRNA,受miR-275调控的Vg-2蛋白为结构不稳定的亲水性酸蛋白,具有良好的抗原性与免疫源性。
【Objective】The aim of the study is to establish the microRNA library of Rhipicephalus microplus,to identify the species of microRNA including the library,to analyze the secondary and tertiary structure of Vg-2 protein regulated by miR-275,and to predict the signal peptides,transmembrane domain,phosphorylation and glycosylation sites,and to dominant epitopes of Vg-2 protein.【Method】The adult female R.microplus was used as sample.The microRNA library of R.microplus was established by using the high-throughput sequencing,and the software miRDP and miRBase database were used to identify the species of microRNA including this library.The online software EXPASY,PRABI and SWISS-MODEL were used to analyze the physical and chemical properties and to infer the secondary and tertiary structure of Vg-2 protein regulated by miR-275.The online software SignalP 5.0,TMHMM,NetPhos 3.1 and NETCGlyc 1.0 were used to detect the signal peptides,transmembrane domain,phosphorylation and glycosylation sites of Vg-2 protein,and the software ABCpred Prediction,Scratch,IEDB and NetCTL were used to predict the dominant antigen epitopes of Vg-2 protein in B and T cells.【Result】Results of our study showed that a microRNA library including total of 383 microRNAs was established,12 of which microRNAs were identified and annotated,371 of which microRNAs were newfound,and 85 microRNAs had the rich contents among these microRNAs.The length of the Vg-2 gene sequence of R.microplus was 5040 bp,and this gene totally encoded 1679 amino acids.Vg-2 protein was a hydrophilic acid protein that was 51.98 kDa in molecular size,and its theoretical isoelectric point(pI)was 6.08.Vg-2 protein usedβsheet as its major structural components.There were 268 phosphate sites and 20 glycosylation sites,but was no signal peptide and transmembrane domain in the amino acids sequence of Vg-2 protein.Vg-2 protein had 58 B cell dominant antigen epitopes and 8 T cell dominant antigen epitopes.【Conclusion】There are 383 microRNAs in the established microRNA lib
作者
李中波
罗世民
杨甜
侯强红
舒鸣
黄翠琴
LI Zhong-bo;LUO Shi-min;YANG Tian;HOU Qiang-hong;SHU Ming;HUANG Cui-qin(Huaihua Vocational and Technical College,Huaihua,Hunan 418000,China;College of Life Science,Longyan University,Longyan,Fujian 364012,China;Engineering Research Center for the Prevention and Control of Animal Original Zoonosis of Fujian Province University,Longyan,Fujian 364012,China)
出处
《西南农业学报》
CSCD
北大核心
2024年第3期678-687,共10页
Southwest China Journal of Agricultural Sciences
基金
湖南省教育厅项目(23B1072)
湖南省自科基金项目(2022JJ50319)
动物源性人兽共患病防控福建省高校工程研究中心项目(2022K004)
怀化市科技局重点项目(2023R2208)。
