用于研究人鼻病毒感染的反向遗传学系统。
Reverse genetics system for studying human rhinovirus infections.
作者信息
Lee Wai-Ming, Wang Wensheng, Bochkov Yury A, Lee Iris
机构信息
Biological Mimetics Inc., 124 Byte Drive, Frederick, MD, 21702, USA,
出版信息
Methods Mol Biol. 2015;1221:149-70. doi: 10.1007/978-1-4939-1571-2_12.
Human rhinovirus (HRV) contains a 7.2 kb messenger-sense RNA genome which is the template for reproducing progeny viruses after it enters the cytoplasm of a host cell. Reverse genetics refers to the regeneration of progeny viruses from an artificial cDNA copy of the RNA genome of an RNA virus. It has been a powerful molecular genetic tool for studying HRV and other RNA viruses because the artificial DNA stage makes it practical to introduce specific mutations into the viral RNA genome. This chapter uses HRV-16 as the model virus to illustrate the strategy and methods for constructing and cloning the artificial cDNA copy of a full-length HRV genome, identifying the infectious cDNA clone isolates, and selecting the most vigorous cDNA clone isolate to serve as the standard parental clone for future molecular genetic study of the virus.
人鼻病毒（HRV）含有一个7.2 kb的正义信使RNA基因组，该基因组进入宿主细胞细胞质后是繁殖子代病毒的模板。反向遗传学是指从RNA病毒RNA基因组的人工cDNA拷贝中再生子代病毒。它一直是研究HRV和其他RNA病毒的强大分子遗传工具，因为人工DNA阶段使得向病毒RNA基因组中引入特定突变成为可能。本章以HRV - 16作为模型病毒，来说明构建和克隆全长HRV基因组的人工cDNA拷贝、鉴定感染性cDNA克隆分离株以及选择最具活力的cDNA克隆分离株作为该病毒未来分子遗传学研究的标准亲本克隆的策略和方法。
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