对虾白斑综合征病毒VP28酵母表面展示  

Yeast surface display of white spot syndrome virus VP28

作　　者：李新新[1] 刘庆慧[1] 张秀丽[1] 黄倢[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《渔业科学进展》2012年第1期61-65,共5页Progress in Fishery Sciences

基　　金：对虾行业专项(200803012);国家自然科学基金(30871942)共同资助

摘　　要：以pYD1为载体,制备以酒精酵母为载体的基因工程免疫制剂。参照GenBank中对虾白斑综合征病毒VP28基因序列设计引物,PCR扩增得到预期长度的产物,双酶切插入用于酒精酵母表面展示的穿梭质粒载体pYD1,转化大肠杆菌TOP10,提取阳性质粒转化酒精酵母菌株EBY100,诱导表达后,用免疫荧光检测外源蛋白的表达。结果获得VP28酵母表面展示菌,测得最佳诱导时间为36～48h。以此展示酵母活细胞分设两个浓度组,腹腔注射螯虾,检测其毒性。结果表明,此重组酵母对螯虾是安全的,该研究为下一步对虾用活载体免疫制剂效果的研究奠定了基础。pYD1 was selected to make a vaccine using Saccharomyces cerevisiae pair as the live carrier. One pair of primers was designed according to the gene encoding white spot syndrome virus （WSSV） of VP28. VP28 gene was obtained by using PCR method. Fragment of VP28 was ligated into pYD1 and then transformed with E. coli TOP10. The construct was propagated in E. coli TOP10 and then was transformed into the yeast strain EBY100. The dis play of VP28 protein on the yeast surface was confirmed by fluorescent staining with antibody a- nalysis. The results showed that VP28 was displayed on yeast surface. The infection test indi cated that the yeast cells displaying VP28 was safe to Procarnbarus clarkii. This work is helpful for further research on the immunological effect of the live vaccine.o

关 键 词：对虾白斑综合征病毒 VP28 酵母表面展示 安全性 

分 类 号：S945.46[农业科学—水产养殖]