轮状病毒Rotateq及Rotarix疫苗VP8~*蛋白的原核表达、纯化与鉴定

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英文篇名：Rotavirus Rotateq and Rotari vaccine VP8~* protein prokaryotic expression, purification and identification 作者：邓中华 ; 郭妮君 ; 陈杰 ; 谭超超 ; 熊艺灿 ; 曹友德 英文作者：Deng Zhong-hua;Guo Ni-jun;Chen Jie;Tan Chao-chao;Xiong Yi-can;Cao You-de;The First Affiliated Hospital of Hunan Normal University, Hunan Provincial People's Hospital;Department of Medical Laboratory, Hunan Provincial Maternal and Child Health Care Hospital; 关键词：轮状病毒 ; Rotateq疫苗 ; Rotarix疫苗 ; 原核表达 ; VP8*蛋白 英文关键词：rotavirus;;rotateq vaccine;;rotari vaccine;;prokaryotic expression;;VP8* protein 中文刊名：HNYG 英文刊名：Journal of Hunan Normal University(Medical Sciences) 机构：湖南师范大学第一附属医院湖南省人民医院;湖南省妇幼保健院; 出版日期：2019-06-17 出版单位：湖南师范大学学报(医学版) 年：2019 期：v.16;No.68 基金：湖南省自然科学基金(2017JJ2151) 语种：中文; 页：HNYG201903002 页数：4 CN：03 ISSN：43-1449/R 分类号：10-13

摘要

目的 :通过大肠埃希菌原核表达体系,分别获得轮状病毒疫苗Rotateq及Rotarix的VP8*基因表达产物,为深入研究轮状病毒VP8*蛋白的结构和功能奠定基础。方法 :基因合成轮状病毒疫苗Rotateq及Rotarix的P[8]型VP8*基因序列,将其克隆到携带有GST标签的pGEX-4T1表达载体中构建重组质粒pGEX-4T1-VP8*,测序正确后转化大肠杆菌BL21(DE3),通过SDS-PAGE电泳检测表达产物,并通过Glutathione Sepharose 4B纯化柱进行纯化,Western Blot鉴定。结果 :SDS-PAGE电泳结果显示分子量52KD处有深染带,Western-Blot显示该处蛋白能与GST标签抗体结合。结论 :成功构建了包含Rotateq及Rotarix VP8*基因的重组表达载体pGEX-4T1-VP8*,获得了可溶性GST-VP8*重组蛋白。
        Objective To obtain Rotavirus vaccine Rotateq and Rotari VP8~* gene expression products by E. coli prokaryotic expression system, further study of rotavirus VP8~* protein structure and function of the foundation. Method Synthesis Rotateq and Rotarix rotavirus vaccine type P [8] VP8~* gene sequences, to clone it to Pgex-4 T1 with GST tags in the expression vector to construct recombinant plasmid, After sequencing confirmed then transformation into E. coli BL21(DE3). SDS-PAGE detection of the expression products, purification by glutathione sepharose 4 B purification column and identification by western blot.Results SDS-PAGE results showed that the molecular weight of 52 KD with hyperchromatic, western blot shows it protein can combine with GST label antibody. Conclusion Successfully built recombinant expression vector Pgex-4 T1-VP8~* which contains Rotateq and Rotarix VP8~* gene, and obtained soluble GST-VP8~* recombinant proteins successful.

引文

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