A chimeric plasmid from cDNA clones of poliovirus and coxsackievirus produces a recombinant virus that is temperature-sensitive.
A chimeric plasmid from cDNA clones of poliovirus and coxsackievirus produces a recombinant virus that is temperature-sensitive.
复制标题
来自脊髓灰质炎病毒和柯萨奇病毒的 cDNA 克隆的嵌合质粒产生了对温度敏感的重组病毒。
DOI：
10.1073/pnas.83.6.1777
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发表时间：
1986
影响因子：
11.1
通讯作者：
Tracy,S
中科院分区：
文献类型：
--
作者：
Semler,BL;Johnson,VH;Tracy,S
我们已经将柯萨奇病毒B3基因组5‘非编码区的405个核苷酸片段插入到脊髓灰质炎病毒RNA基因组的感染性cDNA拷贝中。将含有该杂合基因组结构的质粒DNA导入培养的猴细胞中，产生感染性病毒。重组病毒在37℃下显示出对温度敏感的生长表型。我们发现，与33.5℃相比，在37℃下感染重组病毒的HeLa细胞中病毒蛋白和病毒RNA的水平大幅下降。因此，将柯萨奇病毒基因组的一部分插入脊髓灰质炎病毒基因组中会产生温度敏感的重组病毒。这种替换发生在脊髓灰质炎病毒基因组的一个区域，到目前为止，还没有显示出具有任何编码功能，这表明参与复制酶识别或核糖体结合的RNA序列可能有助于重组病毒的温度敏感表型。
We have inserted a 405-nucleotide fragment from the 5' noncoding region of the coxsackievirus B3 genome into an infectious cDNA copy of the poliovirus RNA genome. Transfection of plasmid DNA containing this hybrid genome construct into cultured monkey cells produced infectious virus. Recombinant virus stocks displayed a temperature-sensitive phenotype for growth at 37 degrees C. We found that there is a dramatic reduction in the level of viral proteins and viral RNAs in HeLa cells infected with the recombinant at 37 degrees C compared to that obtained at 33.5 degrees C. Thus, insertion of a portion of the coxsackievirus genome into the poliovirus genome produces a temperature-sensitive recombinant virus. That this substitution occurs in a region of the poliovirus genome that, to date, has not been shown to have any coding function suggests that RNA sequences involved in replicase recognition or ribosome binding may contribute to the temperature-sensitive phenotype of the recombinant virus.