人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

doi:10.3969/j.issn.1004-616x.2020.01.008

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (1): 43-46,51.doi: 10.3969/j.issn.1004-616x.2020.01.008

人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

徐晓园¹, 李文丽², 徐岚¹, 李蕊²

1. 汕头大学医学院第一附属医院妇产科, 广东汕头 515041;
2. 汕头大学医学院微生物学与免疫学教研室, 广东汕头 515041

收稿日期:2018-11-24 修回日期:2019-10-15 出版日期:2020-01-31 发布日期:2020-02-05
通讯作者: 李蕊,E-mail:07rli1@stu.edu.cn E-mail:07rli1@stu.edu.cn
作者简介:徐晓园,E-mail:g_xyxu@stu.edu.cn。
基金资助:
国家自然科学基金项目（31300761）；广东省自然科学基金项目（2016A030310075）

Expression of VP1 from enterovirus 71 and coxsackievirus A16

XU Xiaoyuan¹, LI Wenli², XU Lan¹, LI Rui²

1. Department of Obstetrics and Gynecology, the First Affiliated Hospital of Shantou University Medical College, Shantou 515041;
2. Department of Microbiology and Immunology, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2018-11-24 Revised:2019-10-15 Online:2020-01-31 Published:2020-02-05

摘要/Abstract

摘要： 目的：克隆人肠道病毒71型（EV71）及柯萨奇病毒A组16型（CA16）的VP1蛋白编码基因，构建相应真核表达质粒在体外进行重组表达，检测其细胞内定位，为EV71及CA16的疫苗研究提供基础。方法：通过PCR扩增分别获取EV71及CA16的VP1编码序列，插入pcFlag载体，分别构建EV71及CA16 VP1与FLAG标签融合的真核表达质粒；瞬时转染人胚肾293T细胞及横纹肌肉瘤RD细胞，Western blot法检测融合蛋白的表达，免疫荧光检测融合蛋白的细胞内定位情况。结果：测序表明两种来源VP1蛋白的重组表达质粒构建正确；分别转染两种不同的细胞后，均可通过Western blot检测到相应蛋白表达；免疫荧光检测显示两种毒株的VP1均表达在细胞质中。结论：外源融合表达EV71或CA16 VP1蛋白的真核表达质粒构建成功；所构建质粒分别转染细胞后，均可在细胞质内检测到VP1融合蛋白的表达。

关键词: 肠道病毒, EV71, 柯萨奇病毒, VP1, 真核表达

Abstract: OBJECTIVE: To clone VP1 genes from enterovirus 71 (EV71) and coxsackievirus A16 (CA16) and to express the genes in eukaryotic cells. METHODS: VP1 coding sequences were obtained by PCR,and then inserted into pcFlag vector to construct FLAG-fused eukaryotic expression plasmids. The recombinant plasmids were used to transfect 293T cells and RD cells,and Western blotting was performed to detect the recombinant proteins. RESULTS: The plasmids containing VP1 were confirmed by sequencing. Recombinant proteins were detected in transfected cells using Western blotting. CONCLUSION: Exogenous expression of plasmids containing VP1 coding sequences of EV71 and CA16 were constructed successfully,and the recombinant proteins was detected in transfected cells.

Key words: enterovirus, EV71, coxsackievirus, VP1, exogenous expression

中图分类号:

Q786

徐晓园, 李文丽, 徐岚, 李蕊. 人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达[J]. 癌变·畸变·突变, 2020, 32(1): 43-46,51.

XU Xiaoyuan, LI Wenli, XU Lan, LI Rui. Expression of VP1 from enterovirus 71 and coxsackievirus A16[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(1): 43-46,51.

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人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

Expression of VP1 from enterovirus 71 and coxsackievirus A16

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