孟少东,姜新宇,葛恒,胡光耀,杨大佐,张家林,李强

• 1:大连海洋大学农业农村部北方海水增养殖重点实验室
• 2:盐城工学院海洋与生物工程学院

摘要(Abstract)：

为进一步研究鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2, CyHV-2)ORF4蛋白的功能,通过PCR扩增CyHV-2 ORF4基因,并构建重组穿梭质粒pFastBac HTA-ORF4,将其转化至大肠杆菌DH10 Bac感受态细胞后获得重组杆粒Bacmid-ORF4;采用脂质体法转染Sf9细胞后,利用间接免疫荧光法(IFA)和Western blot鉴定重组蛋白,采用His亲和层析柱纯化重组蛋白,并通过小鼠免疫试验分析其免疫原性。结果表明:转染72 h后Sf9细胞逐渐呈现出细胞变圆、细胞核明显增大的形态变化;间接免疫荧光结果显示,重组杆状病毒感染的Sf9细胞中出现了明显的绿色荧光;Western blot结果显示,纯化的CyHV-2 ORF4蛋白能够与抗His单抗发生特异性反应;小鼠免疫试验显示,纯化的CyHV-2 ORF4蛋白能诱导小鼠产生特异性免疫反应。研究表明,本研究中获得的CyHV-2 ORF4蛋白有较好的免疫原性,为CyHV-2 ORF4蛋白的功能解析提供了良好材料。

关键词(KeyWords)： 鲤疱疹病毒Ⅱ型(CyHV-2);杆状病毒表达;ORF4蛋白;免疫原性

Abstract:

Keywords:

基金项目(Foundation): 江苏省自然科学基金青年基金(BK20181053);; 江苏省农业科技自主创新资金项目(CX(18)3028);; 盐城工学院人才引进项目(xjr2019044)

作者(Author): 孟少东,姜新宇,葛恒,胡光耀,杨大佐,张家林,李强

DOI: 10.16535/j.cnki.dlhyxb.2020-166

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