【作者】 张炜；

【导师】 雷涵；

【作者基本信息】 西南交通大学 ， 药学， 2019， 硕士

【摘要】 白斑综合症病毒(White Spot Syndrome Virus,WSSV)是虾养殖中最具威胁的传染性病原体之一,其宿主范围广、致死率高、传染性强,给世界虾养殖业造成了巨大的经济损失。遗憾的是,目前尚无有效药物防治,只能通过加强养殖管理措施的方式进行预防。毋庸置疑,研发有效的药物是预防WSSV感染的最有效的措施之一。本论文以WSSV的表面包膜蛋白vp28基因(基因库编号:FJ756456.1)作为研究对象,构建重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28。通过SDS-PAGE、Western blot、免疫荧光分析以及流式细胞仪分析,对重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28的体外表达进行分析。最后,以白肢虾(Penaeus vannamei)作为动物模型,重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28口服给药后,通过WSSV进行攻击实验,检测并分析重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28的保护效率。成功构建的重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28可以稳定表达WSSV的VP28蛋白。WSSV攻击结果显示,白肢虾(Penaeus vannamei)经过口服给药后的存活率为60%。这表明重组酿酒酵母S.cerevisiae EBY100/pYD1-VP28可应用于虾养殖业中预防WSSV的感染,而且还潜藏着巨大的市场应用前景。更多还原

【Abstract】 White spot syndrome virus(WSSV)is one of the most dangerous pathogens infecting shrimp and other crustaceans.It has the characteristics of wide host range,high fatality rate and strong infectivity,which causes great economic losses to the world shrimp breeding industry.Unfortunately,there is no effective drug to prevent WSSV infection,which can only be prevented by strengthening management measures.Undoubtedly,the development of effective drugs is one of the most effective measures to prevent WSSV infection.In this study,recombinant S.cerevisiae EBY100/PYD1-VP28 was constructed based on the envelope protein vp28 gene of WSSV(gene bank number:FJ756456.1)was used as the research object.The expression of S.cerevisiae EBY100/PYD1-VP28 was analyzed by SDS-PAGE,Western blot,immunofluorescence analysis and flow cytometric assay.Then,Penaeus vannamei was used as an animal model and S.cerevisiae EBY100/PYD1-VP28 was orally administrated,the protection efficacy of S.cerevisiae EBY100/PYD1-VP28 was determined by WSSV challenge experiment.S.cerevisiae EBY100/PYD1-VP28 was successfully constructed.WSSV challenge experiment indicated that the survival rate of Penaeus vannamei was 60% after oral administration.This implied that recombinant S.cerevisiae EBY100/PYD1-VP28 could be applied for shrimp aquaculture to prevent WSSV infection,which has a huge market application.更多还原