MOLECULAR-CLONING OF NATURAL PARAMYXOVIRUS COPY-BACK DEFECTIVE INTERFERING RNAS AND THEIR EXPRESSION FROM DNA
MOLECULAR-CLONING OF NATURAL PARAMYXOVIRUS COPY-BACK DEFECTIVE INTERFERING RNAS AND THEIR EXPRESSION FROM DNA
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DOI：
10.1016/0042-6822(92)90166-m
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发表时间：
1992-11-01
期刊：
影响因子：
3.7
通讯作者：
ROUX, L
中科院分区：
文献类型：
--
作者：
CALAIN, P;CURRAN, J;ROUX, L
利用副粘病毒复制回缺陷干扰(DI)RNA的独特序列组织，在不分离其辅助非缺陷基因组的情况下，对仙台病毒(SV)和麻疹病毒(MV)的DI RNA进行了PCR扩增和克隆。克隆的设计是这样的，即T7聚合酶转录的质粒将产生精确的5‘端和3’端的DI RNAs。用痘苗病毒重组体产生的T7聚合酶在BHK细胞中转录出SV DI克隆，并用克隆基因表达的SV-L蛋白、P/C蛋白和NP蛋白进行包装和复制。这样的实验开启了直接检查与病毒增殖有关的顺式作用序列的可能性，而不使用间接标记，如CAT活性。
Using the unique sequence organization of copy-back defective interfering (DI) RNAs of paramyxoviruses, Sendai virus (SV), and measles virus copy-back DI RNAs were PCR amplified and cloned, without having to separate them from their helper nondefective genomes. The cloning was designed so that T7 polymerase transcription of the plasmids would generate DI RNAs with the exact 5′ and 3′ ends. The SV DI clone, transcribed from the plasmid in BHK cells using T7 polymerase produced by a vaccinia virus recombinant, was encapsidated and replicated by the SV-L, P/C, and NP proteins expressed from cloned genes. Such experiments open the possibility of examining thecis-acting sequences involved in viral multiplication directly, without using indirect markers such as CAT activity.