白斑综合症病毒囊膜蛋白VP28单链抗体的酵母表面展示及其抗病毒特性的初步研究

白斑综合症病毒(white spot syndrome virus,WSSV)是对虾养殖业中危害最大的病原微生物之一,对对虾养殖业造成了巨大的经济损失,但目前尚无有效防治手段.本研究从WSSV囊膜蛋白VP28免疫的小鼠细胞中分别扩增到重链可变区基因和轻链可变区基因,大小为351 bp和342 bp.两基因序列通过Linker序列连接后,构建到酵母表面载体pYD1中,得到重组质粒pYD1-vp28scfv.将该重组质粒在酿酒酵母EBY100中诱导表达,通过免疫荧光实验发现,重组蛋白VP28scFv在酿酒酵母EBY100表面实现展示.进一步通过结合实验表明该展示蛋白具有WSSV的结合活性.本研究在结合酿酒酵母可作为饲料添加剂的特性基础上,对WSSV囊膜蛋白VP28单链抗体的抗病毒特性进行了应用,从而为WSSV的防治提供了新思路.

Yeast display of single-chain variable fragment against WSSV envelop protein VP28 and preliminary studies on its antiviral activity

White spot syndrome virus（WSSV）,a strong mortal virus,has caused huge amount of losses in the shrimp aquaculture industry worldwide.However,there are still no control measures for WSSV infection.In this study,a single-chain variable fragment（scFv） against WSSV envelop protein VP28,including heavy（351 bp） and light（342 bp） chain variable regions,was cloned and characterized.Then,this gene-fragment was digested with BamHⅠ/EcoRⅠand ligated into yeast display vector pYD1.Moreover,this construct was transformed into the yeast strain EBY100.Immunofluorescence experiments showed that scFv against VP28 was successfully displayed on the extracellular surface of Saccharomyces cerevisiae cells and had ability to bind to WSSV.Our results provide a potential method for preventing WSSV infection.