建兰花叶病毒广东分离物TGBp1生物信息学分析及抗血清制备  

Bioinformatics analysis and antiserum preparation of Cymbidium mosaic virus TGBp1 of Guangdong isolate

作　　者：宋若楠 饶雪琴[1] SONG Ruonan;RAO Xueqin(Guangdong Province Key Laboratory of Microbial Signals and Disease Control/College of Plant Protection,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]广东省微生物信号与作物病害防控重点实验室/华南农业大学植物保护学院,广东广州510642

出　　处：《河北农业大学学报》2021年第1期97-101,共5页Journal of Hebei Agricultural University

基　　金：广东省现代农业产业技术体系花卉病虫害防治创新团队(2060302).

摘　　要：建兰花叶病毒(Cymbidium mosaic virus,CymMV)是危害兰花的重要病毒。为对建兰花叶病毒广东分离物运动蛋白TGBp1进行研究,本试验制备了TGBp1多克隆抗血清,将CymMV广东分离物TGBp1基因克隆到原核表达载体中,诱导表达融合蛋白,以其为抗原制备多克隆抗血清,并对TGBp1进行生物信息学分析。结果表明,TGBp1融合蛋白的相对分子质量约为31.5 kD,该融合蛋白以包涵体形式存在;并以纯化的融合蛋白为抗原成功制备了TGBp1多克隆抗血清。Western blot分析表明,该抗血清与融合蛋白有较强的免疫反应;间接ELISA结果显示其效价高达524288倍。生物信息学分析表明,该蛋白结构特征与已报道的CymMV其他分离物基本一致。本研究所制备的TGBp1多克隆抗血清具有专化性,为该蛋白质的生物学功能研究奠定了基础。Cymbidium mosaic virus(CymMV)is an important virus that affects orchid’s production.In order to provide a foundation for further research on the function of CymMV TGBp1,a polyclonal antiserum of TGBp1 was prepared in this study.The TGBp1 gene of CymMV isolate was cloned and sequenced,and then inserted into the plasmid pET-28a(+)to construct the prokaryotic expression recombinant plasmid.The fusion protein was induced and purified and analyzed.After that,the purified fusion protein was used as antigen to generate polyclonal antibody.In addition,bioinformatics analysis of TGBp1 was performed by biology software and analysis tools online.The results showed that the expressed fusion protein of His-TGBp1 was about 31.5 kD in size.Soluble analysis indicated that the fusion protein was inclusion body.The titer of the antiserum was 524288 by indirect ELISA,and it had a strong immune response with the His-TGBp1 fusion protein through Western blot.Bioinformatics analysis showed that the structure of TGBp1 was the same as those of other reported CymMV isolates.Briefly,the polyclonal antibody had specificity,which could provide the foundation for the study of biological function of TGBp1.

关 键 词：建兰花叶病毒 运动蛋白 原核表达 抗血清 生物信息学 

分 类 号：S432.41[农业科学—植物病理学]