细粒棘球绦虫泛素结合酶基因家族的生物信息学及表达分析

Bioinformatics and Expression Analysis of Ubiquitin-conjugating Enzyme Gene Family of Echinococcus granulosus

本研究对细粒棘球绦虫泛素结合酶(EgE2s)基因家族进行鉴定、生物信息学分析以及检测其在不同发育阶段的转录水平,为其功能研究和构建优势抗原表位疫苗奠定基础。本研究基于数据库中细粒棘球绦虫基因组学信息,对EgE2s基因家族进行T-A克隆、生物信息学分析以及采用qRT-PCR法检测了这些基因在原头蚴(Protoscolex, PSCs)和28日龄童虫中的转录水平。结果成功克隆并测序19个EgE2s基因,更正了两个原序列有误的EgE2s,分别为EgE2L3(登录号:MZ277618);EgE2J1(登录号:MZ277619)。三级结构和保守基序预测结果表明EgE2s高度保守。qRT-PCR结果显示,EgE2s的转录水平在PSCs和28日龄童虫中存在差异,其中EgE2A(P<0.001)、EgE2J1(P<0.01)在PSCs阶段高表达。B细胞抗原表位预测表明EgE2N的139-157区域位于EgE2s保守基序外。同时,T细胞抗原表位预测表明EgE2N与人和犬MHCⅠ类分子各有两个强结合位点,且有一个共同的抗原表位位点104-113。综上,EgE2A和EgE2J1可能在PSCs的生长发育中起重要作用。EgE2N的139-157和104-113区域可能是药物研究和疫苗开发的靶序列。

To further explore the gene function and construct the dominant epitope vaccine,sequences identification,bioinformatics and transcription analysis of ubiquitin-conjugating enzyme gene family of Echinococcus granulosus sensu lato(EgE 2s)were conducted.Based on the genomic data of Echinococcus granulosus published before,the EgE 2s gene family was cloned and analyzed by bioinformatics.In addition,the transcriptional levels of EgE 2s in protoscolex(PSCs)and 28-day strobilated worms were detected by qRT-PCR.The results showed that,nineteen 19 EgE 2s genes were cloned and sequenced successfully.Among them,two CDS sequences among two of them were originally wrong had been corrected and were submitted to NCBI with to obtain accession number(EgE 2L3:MZ277618;EgE 2J1:MZ277619).The prediction of tertiary structures and conservative motifs indicated that EgE 2s were highly conserved.qRT-PCR showed that the transcriptional levels of EgE 2s in PSCs differed from 28-day strobilated worms,EgE 2A(P<0.001)and EgE 2J1(P<0.01)were highly expressed in PSCs.The prediction of B cell epitopes showed that the 139-157 region of Eg E2N was located outside the conserved motifs of Eg E2s.Meanwhile,the T cell epitopes prediction showed that Eg E2N had two strong binding sites with human and canine MHC I molecules,respectively,including a common epitope region 104-113.In conclusion,EgE 2A and EgE 2J1 may play important roles in the growth and development of PSCs.The 139-157 and 104-113 regions of Eg E2N may be the target sequences for the development of drugs and vaccines.