IDENTIFICATION OF A DOMAIN WITHIN THE PHOSPHOPROTEIN OF VESICULAR STOMATITIS-VIRUS THAT IS ESSENTIAL FOR TRANSCRIPTION INVITRO
IDENTIFICATION OF A DOMAIN WITHIN THE PHOSPHOPROTEIN OF VESICULAR STOMATITIS-VIRUS THAT IS ESSENTIAL FOR TRANSCRIPTION INVITRO
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DOI：
10.1073/pnas.83.23.8873
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发表时间：
1986-12-01
影响因子：
11.1
通讯作者：
BANERJEE, AK
中科院分区：
文献类型：
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作者：
GILL, DS;CHATTOPADHYAY, D;BANERJEE, AK
将水泡性口炎病毒（新泽西血清型）磷蛋白（NS）mRNA的全长cDNA拷贝插入pGEM 4载体噬菌体SP 6 RNA聚合酶启动子下游。在体外转录的cDNA导致在NS mRNA的合成，随后在无细胞兔网织红细胞系统中翻译成NS蛋白。以纯化的病毒L蛋白和N-RNA为模板，体外转录重组合成mRNA，证明表达的NS蛋白具有生物学活性。NS基因的缺失定位在氨基酸残基213和247之间定义了一个特定的结构域，这是体外转录所必需的。另一方面，去除COOH末端的21个氨基酸对转录没有显著影响。该结构域似乎参与NS蛋白与N蛋白-RNA模板的有效结合。
A full-length cDNA copy of the phosphoprotein (NS) mRNA of vesicular stomatitis virus (New Jersey serotype) was inserted into pGEM4 vector downstream of the promoter for bacteriophage SP6 RNA polymerase. Transcription of the cDNA in vitro resulted in the synthesis of NS mRNA, which was subsequently translated into NS protein in a cell-free rabbit reticulocyte system. The biological activity of the expressed NS protein was demonstrated by in vitro synthesis of mRNA by transcription-reconstitution with purified viral L protein and N-RNA template. Deletion mapping of the NS gene defined a specific domain between amino acid residues 213 and 247, which was essential for in vitro transcription. Removal of the COOH-terminal 21 amino acids, on the other hand, did not have a significant effect on transcription. This domain appears to be involved in efficient binding of NS protein to the N protein-RNA template.