鸭坦布苏病毒E蛋白重组乳酸菌的构建与鉴定
Expression and Characterization of Tembusu Virus E Protein in Recombinant Lactic Acid Bacteria
张弛 1高玉宸 1杨晶 1王林 2张朕鹏 3刁有江1
作者信息
- 1. 山东农业工程学院,山东 济南 250100
- 2. 济南亿民动物药业有限公司,山东 济南 250100
- 3. 山东鲁港福友药业有限公司,山东 济南 250101
- 折叠
摘要
E蛋白为鸭坦布苏病毒(Tembusu virus,TMUV)表面的囊膜蛋白,是该病毒主要的抗原蛋白.为构建表达鸭坦布苏病毒E蛋白的重组乳酸菌,本研究首先通过比对GenBank中已发布的DTMUV E基因序列,扩增E基因全长并纯化,构建pMD19-T-E重组克隆载体;以pMD19-T-E重组载体为基础,利用同源重组技术将E基因重组至乳酸菌载体pNZ8149,菌液PCR及测序表明,本研究成功构建了pNZ8149-TMUV-E重组乳酸菌表达载体.SDS-PAGE试验结果显示重组乳酸菌pNZ8149-TMUV-E可成功表达E蛋白.综上所述,本研究成功构建了能够表达鸭坦布苏病毒E蛋白的重组乳酸菌,为进一步开发鸭坦布苏病毒病的新型疫苗奠定了基础.
Abstract
The E protein is an envelope protein located on the surface of the duck Tembusu virus(TMUV)and serves as its primary antigenic protein.To construct recombinant lactic acid bacteria(LAB)expressing the TMUV E protein,this study first amplified and purified the full-length E gene by comparing it with the DTMUV E gene sequences available in GenBank.A recombinant cloning vector,pMD19-T-E,was successfully constructed.Using the pMD19-T-E recombinant vector as a template,the pNZ8149-TMUV-E/NZ3900 LAB expression vector was developed through homologous recombination.Both bacterial colony PCR and sequencing confirmed the successful construction of the pNZ8149-TMUV-E recombinant LAB expression vector.SDS-PAGE analyses further demonstrated that the recombinant LAB pNZ8149-TMUV-E efficiently expressed the E protein.In conclusion,this study successfully engineered recombinant LAB capable of expressing the duck Tembusu virus E protein,laying a foundation for the future development of novel vaccines against TMUV infection.
关键词
鸭坦布苏病毒/E蛋白/乳酸菌Key words
Duck tembusu virus/E protein/Lactic acid bacteria引用本文复制引用
出版年
2025
NETL
NSTL
万方数据