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首页> 外文期刊>Journal of Virology >Construction of a specific amber codon in the simian virus 40 T-antigen gene by site-directed mutagenesis.
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Construction of a specific amber codon in the simian virus 40 T-antigen gene by site-directed mutagenesis.

机译：通过点定向诱变在Simian病毒40 T-antigen基因中构建特异琥珀墓穴。

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The site-directed bisulfite mutagenesis technique has been used to construct a specific mutation, am404, at nucleotide position 3124 in the simian virus 40 genome. The mutation was contained within a PstI restriction site (map position 0.27) and prevented cleavage by PstI at that position. Nucleotide sequence analysis of the mutagenized region indicated that only a single base pair change had occurred: a guanosine x cytosine leads to adenine x thymine transition. Comparison of the nucleotide sequence of am404 with the known DNA sequence of simian virus 40 indicted that the mutation in am404 resulted in the conversion of a glutamine codon to an amber codon. am404 could not replicate autonomously when transfected into monkey cells (BSC-40) but did replicate when it was cotransfected with the late deletion helper virus dl1007. On the basis of its position in the T-antigen, gene am404 should produce a T-antigen 24% shorter than the wild-type protein.
机译：已经使用位点定向的亚硫酸氢盐诱变技术在Simian病毒40基因组中构建特定突变，AM404，核苷酸位置3124。突变含有PSTI限制性位点（MAP位置0.27）并通过该位置的PSTI防止切割。诱变区的核苷酸序列分析表明，仅发生单一碱基对变化：鸟氨酸X细胞苷导致腺嘌呤X胸腺嘧啶转变。 AM404核苷酸序列与已知的DNA序列与Simian病毒40的已知DNA序列的比较引起了AM404中的突变导致谷氨酰胺密码子转化为琥珀密码子。在转染到猴细胞（BSC-40）转染时，am404无法自主复制，但是当用晚期删除辅助病毒DL1007进行COTANSFECTE时恢复。基于其在T-antigen中的位置，基因AM404应产生比野生型蛋白质短的T-抗原24％。

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