| 李新新,刘庆慧,张秀丽,黄倢.对虾白斑综合征病毒VP28酵母表面展示.渔业科学进展,2012,33(1):61-65 |
| 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 |
| 投稿时间：2011-02-19 修订日期：2011-10-30 |
| DOI： |
| 中文关键词: 对虾白斑综合征病毒 VP28 酵母表面展示 安全性 |
| 英文关键词: WSSV VP28 Yeast surface display Safety |
| 基金项目:对虾行业专项（200803012）和国家自然科学基金（30871942）共同资助 |
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| 中文摘要: |
| 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36～48 h。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。 |
| 英文摘要: |
| 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 display of VP28 protein on the yeast surface was confirmed by fluorescent staining with antibody analysis. The results showed that VP28 was displayed on yeast surface. The infection test indicated that the yeast cells displaying VP28 was safe to Procambarus clarkii. This work is helpful for further research on the immunological effect of the live vaccine. |
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