发热伴血小板减少综合征病毒蛋白Gc原核表达及免疫原性研究

doi:10.11853/j.issn.1003.8280.2019.04.006

中国媒介生物学及控制杂志 ›› 2019, Vol. 30 ›› Issue (4): 383-386.DOI: 10.11853/j.issn.1003.8280.2019.04.006

发热伴血小板减少综合征病毒蛋白Gc原核表达及免疫原性研究

姚文武, 颜浩, 楼秀玉, 潘军航, 孙逸, 茅海燕, 张严峻

浙江省疾病预防控制中心微生物检验所, 浙江杭州 310051

收稿日期:2019-03-04 出版日期:2019-08-20 发布日期:2019-08-20
通讯作者: 张严峻,Email:yjzhang@cdc.zj.cn
作者简介:姚文武,男,技师,主要从事微生物检验工作,Email:wwyao@cdc.zj.cn
基金资助:
浙江省医药卫生科技项目（2018KY341，2017KY035，2017KY290）；国家科技重大专项（2018ZX10734401）

Prokaryotic expression and immunogenicity of severe fever with thrombocytopenia syndrome virus glycoprotein Gc

YAO Wen-wu, YAN Hao, LOU Xiu-yu, PAN Jun-hang, SUN Yi, MAO Hai-yan, ZHANG Yan-jun

Zhejiang Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang Province, China

Received:2019-03-04 Online:2019-08-20 Published:2019-08-20
Supported by:
Supported by the Health Care General Studies Program in Zhejiang Province (No. 2018KY341,2017KY035,2017KY290) and National Science and Technology Major Project of China (No. 2018ZX10734401)

摘要/Abstract

摘要： 目的对原核表达发热伴血小板减少综合征病毒（SFTSV）蛋白Gc免疫原性进行初步研究。方法参照SFTSV HB29病毒株包膜蛋白Gc编码基因，通过大肠埃希菌密码子偏好性优化后化学合成方法得到Gc目的基因。将目的基因导入到载体pET-His中，得到表达载体pET-His-Gc。经测序确认后转化到BL21（DE3）感受态细胞中表达，纯化后进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western blot验证。蛋白免疫新西兰大白兔后收集兔血清进行抗体效价测定。结果成功构建了pET-His-Gc融合蛋白表达载体，并在大肠埃希菌中得到可溶性表达，且得到的Gc蛋白具有良好的免疫原性。动物实验结果表明，3次免疫后兔血清的抗体效价可达到1 ∶ 512 000。结论实现了SFTSV包膜蛋白Gc的原核表达，为后续SFTSV包膜蛋白的研究奠定了基础。

关键词: 发热伴血小板减少综合征病毒, 原核表达, 免疫原性

Abstract: Objective To express severe fever with thrombocytopenia syndrome virus (SFTSV) glycoprotein Gc in prokaryotic cells, and to preliminarily study the immunogenicity of glycoprotein Gc. Methods The DNA sequence encoding envelope glycoprotein Gc in SFTSV HB29 strain was chemically synthesized after codon optimization based on Escherichia coli codon preference. The expression vector pET-His-Gc was constructed by cloning the target gene into the vector pET-His. The sequence of the vector was confirmed by sequencing. Protein expression was carried out by transforming the vector into BL21 (DE3) competent cells. Target protein was purified and verified by SDS-PAGE and Western blot. Purified protein was used to immunize New Zealand white rabbits. The rabbit sera were collected for determination of antibody titer. Results The expression vector pET-His-Gc was successful constructed. Glycoprotein Gc was expressed in E. coli as a soluble protein. The obtained Gc protein showed good immunogenicity. Animal experiments showed that the rabbit serum antibody titer reached 1:512 000 after three immunizations. Conclusion In this study, SFTSV envelope glycoprotein Gc is successfully expressed in prokaryotic cells, which paves the way for subsequent studies of SFTSV envelope proteins.

Key words: Severe fever with thrombocytopenia syndrome virus, Prokaryotic expression, Immunogenicity

中图分类号:

R373.3

姚文武, 颜浩, 楼秀玉, 潘军航, 孙逸, 茅海燕, 张严峻. 发热伴血小板减少综合征病毒蛋白Gc原核表达及免疫原性研究[J]. 中国媒介生物学及控制杂志, 2019, 30(4): 383-386.

YAO Wen-wu, YAN Hao, LOU Xiu-yu, PAN Jun-hang, SUN Yi, MAO Hai-yan, ZHANG Yan-jun. Prokaryotic expression and immunogenicity of severe fever with thrombocytopenia syndrome virus glycoprotein Gc[J]. Chines Journal of Vector Biology and Control, 2019, 30(4): 383-386.

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发热伴血小板减少综合征病毒蛋白Gc原核表达及免疫原性研究

Prokaryotic expression and immunogenicity of severe fever with thrombocytopenia syndrome virus glycoprotein Gc

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