鸡卵黄抗体技术在5种草莓病毒快速检测中的应用  

Application of chicken yolk antibody technique in rapid detection of five strawberry viruses

作　　者：杨菊梅 何永红 马建忠[3] 周延峰 周小春[1] YANG Ju-mei;HE Yong-hong;MA Jian-zhong;ZHOU Yan-feng;ZHOU Xiao-chun(Cuiying Biomedical Research Center,The Second Hospital of Lanzhou University,Lanzhou 730030,China;Huining County Market Supervision and Administration Bureau,Baiying 730799,Gansu,China;School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China)

机构地区：[1]兰州大学第二医院萃英生物医药研究中心,兰州730030 [2]会宁县市场监督管理局,甘肃白银730799 [3]兰州理工大学生命科学与工程学院,兰州730050

出　　处：《兰州大学学报（自然科学版）》2021年第3期353-359,368,共8页Journal of Lanzhou University(Natural Sciences)

基　　金：兰州市科技计划项目(2014-RC-64)。

摘　　要：利用生物信息学技术对草莓轻型黄边病毒(SMYEV)、草莓斑驳病毒(SMV)及草莓皱缩病毒(SCV)等5种病毒抗原表位进行预测,将预测的抗原肽段及串联表位肽段的DNA片段在大肠埃希氏菌(Escherichia)BL21(DE3)进行原核表达,并将纯化后的抗原表位肽免疫京红一号产蛋鸡.从免疫蛋鸡卵黄中分离卵黄抗体(IgY),并对其进行抗体活性、效价及灵敏度的分析.结果表明,预测到的优势抗原表位肽段分别位于5种草莓病毒外壳蛋白氨基酸序列的27-38、49-82、288-314、62-75、218-238位.5种优势抗原表位肽及其串联抗原表位肽(5S)均能在Escherichia BL21(DE3)中成功表达.5S免疫蛋鸡产生的IgY对于5种草莓病毒均具有较强的免疫活性,抗体滴度为1∶1600,检测灵敏度为10ng.本研究制备的串联抗原表位重组蛋白能诱导蛋鸡产生具有较强免疫活性的IgY抗体,可用于SMYEV、SMV及SCV等5种草莓病毒的同时检测.The antigenic epitopes of strawberry mild yellow edge virus(SMYEV),strawberry mottle virus(SMV)and strawberry crinkle virus(SCV)were predicted with the bioinformatics technology.The predicted DNA fragment of antigenic peptide and tandem epitope were expressed in Escherichia BL21(DE3),and Jinghong No.1 laying hens were immunized with products of purified epitope peptides.The yolk antibody(IgY)was isolated from the yolk and the activity,titer and sensitivity of the antibody IgY were analyzed.The results showed that the peptides of the dominant epitopes were located at 27-38,49-82,288-314,62-75 and 218-238 of the amino acid sequences of the coat proteins of the five strawberry viruses.Five dominant epitope peptides and their tandem epitope peptides(5 S)were successfully expressed in Escherichia BL21(DE3).The IgY produced by layers immunized with 5 S tandem epitope peptide had a strong immune activity against the five strawberry viruses,the antibody titer was 1:1600 and the detection sensitivity was 10 ng.The recombinant protein with the tandem epitope could induce the layers to produce the Ig Y antibody with a strong immune activity,which might be used to detect SMYEV,SMV,SCV,etc simultaneously.

关 键 词：草莓病毒 外壳蛋白 抗原表位 原核表达 鸡卵黄抗体 

分 类 号：Q812[生物学—生物工程]