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当前位置: 首页 > 文章 > 虎源猫泛白细胞减少症病毒VP2基因主要抗原表位区的原核表达和蛋白纯化 东北林业大学学报 2010,38 (6) 101-104
Position: Home > Articles > Prokaryotic Expression of VP2 Antigenic Epitope Gene of Feline Panleukopenia Virus and Its Purification Journal of Northeast Forestry University 2010,38 (6) 101-104

虎源猫泛白细胞减少症病毒VP2基因主要抗原表位区的原核表达和蛋白纯化

作  者：
田丽红;华育平
单  位：
东北林业大学
关键词：
猫泛白细胞减少症病毒;VP2基因;抗原表位;原核表达;蛋白纯化;虎
摘  要：
利用PCR技术从FPV-HLJ株病毒细胞培养物中扩增VP2基因主要抗原表位区片段VP2’。扩增片段克隆至pMD18-T载体,经核苷酸序列测定确认后,亚克隆于pGEX-6p-1原核表达载体,构建pGEX-6p-VP2’重组原核表达质粒。将该质粒转化至表达菌BL21(DE3)中用IPTG进行诱导表达。表达蛋白采用切胶法纯化,并用SDS-PAGE和Westernblot检测纯化蛋白纯度和抗原特异性。结果表明,VP2基因全长1200bp,编码400个氨基酸。重组菌可表达相对分子量约为66kD的融合蛋白,包括目的蛋白40kD和GST标签26kD。该蛋白以包涵体形式存在,且GST融合蛋白具有良好抗原特异性。
译  名：
Prokaryotic Expression of VP2 Antigenic Epitope Gene of Feline Panleukopenia Virus and Its Purification
作  者：
Tian Lihong,Hua Yuping( School of Wildlife Resources,Northeast Forestry University,Harbin 150040,P. R. China)
关键词：
Feline panleukopenia virus; VP2 gene; Antigenic epitope; Prokaryotic expression; Protein purification; Tigers
摘  要：
The gene encoding antigenic epitope VP2 was amplified from feline panleukopenia virus ( FPV) FPV-HLJ strain isolated from tiger feces. The amplified DNA was cloned into the prokaryotic expression vector pGEX-6p-1 after its sequence was confirmed by sequencing. The expression vector was then transferred into E. coli BL21( DE3) for expression under IPTG treatment. The recombinant protein was isolated using SDS-PAGE and recovered from the gel. A second SDS-PAGE was performed to confirm its purity and detect its antigenic specificity by Western blotting. Results indicated that VP2 gene of FPVHLJ was 1 200 bp in length encoding 400 amino acids. A fusion protein of about 66 kD was obtained from the prokaryotic expression system,including 40 kD VP2 protein and 26 kD GST-tag protein. The VP2 protein had high antigenic specificity.

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