Isolation of mutants of an animal virus in bacteria.
Isolation of mutants of an animal virus in bacteria.
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细菌中动物病毒突变体的分离。
DOI：
10.1126/science.6251547
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发表时间：
1980
期刊：
影响因子：
56.9
通讯作者：
D. Nathans
中科院分区：
文献类型：
--
作者：
K. Peden;J. Pipas;S. Pearson;D. Nathans
动物病毒的突变体可以通过重组DNA方法在细菌中分离。由于重组体在细菌中繁殖不需要病毒功能，因此可以分离出顺式或反式作用元件发生致死性变化的病毒突变体，以及部分或条件缺陷突变体。在具有小DNA基因组的病毒的情况下，例如致瘤性猿猴病毒40（SV40），可以将整个病毒DNA插入细菌质粒pBR 322中并在大肠杆菌中克隆。含有单拷贝SV40 DNA的重组质粒在培养物中引起小鼠细胞的形态转化，其效率与从病毒感染的猴细胞中分离的SV40 DNA相同，但重组DNA是非感染性的，并且在允许细胞中复制较差。然而，从质粒上切下的SV40 DNA与真实的病毒DNA一样复制，并且是完全感染性的。通过重组DNA的体外位点特异性或随机局部诱变，然后在E.杆菌许多这样分离的突变体在特定的病毒功能上是有缺陷的。
Mutants of animal viruses can be isolated in bacteria by recombinant DNA methods. Since no viral functions are required for propagation of recombinants in bacteria, viral mutants with lethal changes in cis- or trans-acting elements can be isolated, as well as partially or conditionally defective mutants. In the cases of viruses with small DNA genomes, such as the tumorigenic simian virus 40 (SV40), the entire viral DNA can be inserted into the bacterial plasmid pBR322 and cloned in Escherichia coli. Recombinant plasmids with a single copy of SV40 DNA cause morphological transformation of mouse cells in culture with the same efficiency as SV40 DNA isolated from virus-infected monkey cells, but the recombinant DNA is noninfectious and replicates poorly in permissive cells. However, SV40 DNA excised from the plasmid replicates as well as authentic viral DNA and is fully infectious. SV40 mutants with small deletions or base substitutions have been isolated by in vitro site-specific or random local mutagenesis of recombinant DNA followed by cloning in E. coli. Many of the mutants thus isolated are defective in specific viral functions.