戊型肝炎病毒基因3型衣壳蛋白与Huh-7细胞互作蛋白的筛选  

Screening of interactionbetween capsid protein type 3 of hepatitis E virus gene and Huh-7 cells

作　　者：梁鹏[1,2] 宋阳冉 连瑞雅 陈轶霞 李慧霞[1] Liang Peng;Song Yangran;Lian Ruiya;Chen Yixia;Li Huixia(State Ethnic Affairs Commission Key Laboratory of Biological Engineering and Technology,Biomedical Research Center,Northwest Minzu University,Gansu Lanzhou 730030;College of Life Science and Engineering,Northwest Minzu University,Gansu Lanzhou 730030)

机构地区：[1]西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,甘肃兰州730030 [2]西北民族大学生命科学与工程学院,甘肃兰州730030

出　　处：《现代畜牧兽医》2023年第10期27-31,共5页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：西北民族大学引进人才科研项目(xbmuyjrc2020021);甘肃省青年科技基金计划(21JR1RA212);2022年甘肃省优秀研究生“创新之星”项目(2022CXZX-207)。

摘　　要：试验旨在筛选与戊型肝炎病毒基因3型(HEV)衣壳蛋白(ORF2)互作的宿主蛋白,构建ORF2的真核表达质粒,转染肝癌细胞(Huh-7)后进行免疫共沉淀(Co-IP)试验,将产物进行蛋白质谱分析,筛选与ORF2互作的细胞蛋白,并进行生物信息学分析。结果显示,重组质粒pCAGEN-Myc-ORF2在Huh-7中正确表达,据蛋白质谱分析获得了与ORF2互作的37个候选蛋白;生物信息学分析发现,这些蛋白主要参与RNA剪接的调控、细胞质翻译等生物过程,同时也形式参与如钙黏蛋白结合、泛素-蛋白质连接酶等功能,并且影响核糖体合成途径、参与新型冠状病毒感染等。The aim of the experiment was to screen the host protein interacting with the HEV ORF2,construct the eukaryotic expression plasmid of ORF2,transfect liver cancer cells(Huh-7)and Co-IP test.The product was analyzed by protein spectrum,and the cell proteins interacting with ORF2 were screened.Bioinformatics analysis was performed.The results showed that the recombinant plasmid pCAGEN-Myc-ORF2 was correctly expressed in Huh-7,and 37 candidate proteins interacting with ORF2 were obtained by protein spectrum analysis.Bioinformatics analysis found that these proteins mainly participate in the regulation of RNA splicing,cytoplasmic translation and other biological processes,but also participate in functions such as cadhrin binding,ubiquitin-protein ligase and other functions,and affect ribosome synthesis pathway and participate in novel coronavirus infection.

关 键 词：戊型肝炎病毒 衣壳蛋白 蛋白质谱分析 互作蛋白 生物信息学分析 

分 类 号：S855[农业科学—临床兽医学]