来自cDNA克隆的转录中VSV的感染性缺陷干扰颗粒
A K Pattnaik1, L A Ball, A W LeGrone
1Department of Microbiology, University of Alabama Medical School, Birmingham 35294.
Cell
|June 12, 1992
概括
研究人员从cDNA克隆中产生了囊性口腔炎病毒 (VSV) 的传染性缺陷干扰 (DI) 颗粒. 这种新的方法绕过了对助手VSV的需求,使得有效的病毒颗粒生产和复制研究成为可能.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 缺陷干扰 (DI) 颗粒对于理解病毒复制和进化至关重要.
- 以前生成DI颗粒的方法需要辅助病毒,限制了实验控制.
- 膀性口腔炎病毒 (VSV) 是研究负链RNA病毒的一个模型系统.
研究的目的:
- 开发一种方法,完全从cDNA克隆中生成VSV的传染性DI颗粒.
- 为了证明这种方法不需要辅助VSV.
- 建立一个高水平复制和生物化学分析DI颗粒的系统.
主要方法:
- 从VSV DIRNAcDNA克隆中转录一个完整的负链基因组RNA,使用菌体T7RNA聚合酶.
- 从宿主细胞中单独的cDNA克隆中共同表达五种VSV蛋白质.
- 使用型肝炎病毒 ribozyme 精确的 3' 端生成的转录.
- 在没有辅助病毒的情况下,DI颗粒的封装,复制,组装和芽.
主要成果:
- 仅从cDNA克隆中成功生成VSV的传染性DI颗粒.
- 在不需要辅助VSV的情况下,证明了DI粒子的高水平复制.
- 证实了精确的3'终端对于有效复制的关键作用.
结论:
- 这项研究提出了一种新的,独立于助手的方法,用于从cDNA产生传染性VSV DI颗粒.
- 开发的系统允许对DI粒子复制和组装进行受控研究.
- 这种方法对病毒学研究,疫苗开发和基因治疗载体设计有重大影响.
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