新城疫病毒F蛋白结构域基因原核表达与抗原表位分析

被引：5

作者：

张桂芝

陈亚波

徐程

张朝政

方维焕

机构：

[1] 浙江大学动物预防医学研究所浙江省动物预防医学重点实验室

来源：

浙江大学学报(农业与生命科学版) | 2006年 / 01期

基金：

浙江省自然科学基金;

关键词：

新城疫病毒; 融合蛋白; 结构域; 抗原表位; 原核表达;

D O I：

暂无

中图分类号：

S852.65 [家畜病毒学];

学科分类号：

摘要：

以含新城疫病毒(NDV)融合蛋白(F)基因的重组质粒为模板,设计特异引物进行重叠-延伸PCR扩增获得F基因的抗原结构域基因片段Fa、Fb和Fa-l-Fb,经SalⅠ和NotⅠ酶切后,定向插入原核表达载体pGEX-6P-1,获得重组质粒pGEX-Fa、pGEX-Fb和pGEX-Fa-l-Fb;用定点突变方法获得重组质粒pGEX-Fa-mut.将重组质粒转化大肠杆菌BL21,转化菌经IPTG诱导、提取表达产物进行蛋白电泳和免疫印迹分析.结果表明:Fa-mut、Fb和Fa-l-Fb结构域基因均获得了融合表达,表达产物与NDV阳性血清具有免疫反应性;与pGEX-Fa-mut不同,pGEX-Fa重组质粒在大肠杆菌中未能表达,且转化菌在IPTG诱导过程中未见生长,提示未经突变的Fa片段可能为毒性蛋白.三维结构预测结果显示,目的蛋白片段中均保留F蛋白中原有相应的抗原表位.

引用

页码：82 / 87

页数：6

共 15 条

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