【摘要】 以马立克氏病病毒(MDV)特超强毒648株基因组DNA为模板,通过聚合酶链式反应(PCR)方法扩增出gE基因。将其插入到表达性质粒pGEX-6p-1中谷胱甘肽转移酶(GST)基因的下游,转化大肠杆菌,经筛选、鉴定获得重组质粒,测序结果证明重组质粒的阅读框架不变,将其命名为pG648E。经SDS-PAGE电泳及Westernblotting分析,在82ku有GST-gE的蛋白带,重组质粒在大肠杆菌BL21中以融合蛋白的形式表达。结果表明:MDVgE基因原核表达成功。更多还原

【Abstract】 Marek′s disease virus (MDV) glycoprotein E (gE) gene was amplificated from CEF genomic DNA infected with MDV 648 strain by polymerase chain reaction (PCR). PCR products of the gE gene was inserted into the downstream of GST gene in pGEX6p1 vector. Restriction endonucleases analysis was used to identify the recombinant plasmid pG648E and sequencing by comparing with the sequence of MDV648 strain gE gene. The recombinant plasmid kept the right open reading frame (ORF) and was transformed into host Escherichia coli BL21 for expression. The expression of 648 strains gE gene was confirmed by SDSPAGE and Western blotting with specific antibody, and found a 82 ku in size as protein with GST. The result showed that recombinant plasmid expressed the gE gene fusion protein of MDV.更多还原