重组猪三角型冠状病毒的工程工程稳定表达外基因基因
Meng Yuan1, Yuhan Zhang1, Longfei Chen1
1National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, the Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
猪三角冠状病毒 (PDCoV) 逆遗传系统被开发出来. 用EGFP取代NS6基因创造了一个稳定的PDCoV疫苗载体,使得外来基因表达成为可能.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 猪三角冠状病毒 (PDCoV) 由于其免疫性和宿主热带性,显示出作为疫苗载体的潜力.
- 了解PDCoV的转录调节序列 (TRS) 和遗传稳定性对于载体的发展至关重要.
研究的目的:
- 为了建立一个PDCoV反向遗传系统.
- 为了评估PDCoV TRS元素的转录活性.
- 为了产生一个稳定的,外来基因表达的PDCoV载体.
主要方法:
- 建立了PDCoV菌株DHeB1.1的逆遗传系统.
- 评估了由六个不同的TRS元素驱动的EGFP表达.
- 通过将EGFP插入N-3'基因UTR区域来生成复合病毒.
- 用EGFP取代NS6基因以增强遗传稳定性.
主要成果:
- 所有测试的TRS元素都启动了EGFP表达,NS6 TRS显示了最高的活动.
- 在N-3' UTR基因中与EGFP重组病毒是基因不稳定的.
- 用EGFP取代NS6基因导致稳定的重组病毒传播超过15个通道.
- 稳定的重组病毒保持了一致的EGFP表达.
结论:
- 为PDCoV建立了一个功能性的逆转基因平台.
- PDCoV TRS 元素表现出不同的转录活动.
- 用外来基因替换辅助NS6基因为基于PDCoV的稳定载体发展提供了一种战略.
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