摘要
反向遗传系统可以对RNA病毒直接进行遗传操作 ,为RNA病毒的分子生物学研究提供了一种强大的工具。在过去 2 0年 ,特别是自 90年代中期第一例负链RNA病毒感染性克隆构建成功以来 ,动物RNA病毒的分子生物学研究取得了长足的进展 ,这很大程度上归功于各种动物RNA病毒反向遗传系统的建立。这里系统总结了人类及动物非反转录RNA病毒中各类代表性成员在建立反向遗传系统时的方案设计、遇到的困难及研究者如何克服这些困难。分类讨论到的代表性病毒种属有脊髓灰质炎病毒、冠状病毒 (包括SARS病毒 )、黄病毒、野田村病毒、流感病毒、传染性法氏囊病病毒以及呼肠孤病毒等。
The recovery of the virus from genetic materials in in vitro culture systems or sensitive animals is called virus rescue. A functional infectious clone of RNA virus provides unlimited possibility for genetic studies and the related reverse genetics system that allows directed genetic manipulation of an RNA virus is an extremely powerful research tool. In the past twenty years, especially since the first infectious clone of a negative-stranded RNA virus was reported in the mid-1990's, the reverse genetics systems have been available for nearly all the major human and animal RNA virus groups. The article reviews the progress of this technology, highlighting the obstacles in the construction of reverse genetics systems for major groups of human as well as animal RNA viruses and how the virologists overcame them. There are mainly four external expression systems for construction of the RNA virus reverse genetics systems basing on the kind of RNA viruses. These systems include in vitro RNA transcripts, RNA polymeraseⅠ-driven expression plasmids, RNA polymeraseⅡ-driven expression plasmids and modified vaccinia virus/T7 RNA polymerase-driven expression system. In particular, the viral nucleoprotein and polymerase proteins are required to assemble the viral ribonucleoprotein(RNP) complexes for the rescue of the negative-stranded RNA viruses. Relevant topics about the rescue of the typical viruses are discussed, including poliovirus with the de novo synthesis, Coronaviridae with the largest size of genome, Flaviviridae with the instable clones, HCV with the quasispecies nature, nodaviruses with the virus-host interaction, influenza virus with the RNA polⅠ transcription system, Arenavirdae with the ambisense coding strategies etc.
出处
《生物工程学报》
CAS
CSCD
北大核心
2004年第3期311-318,共8页
Chinese Journal of Biotechnology
基金
浙江省重点科技项目基金资助 (No .2 0 0 3C2 2 0 0 2 )~~
