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首页> 外文期刊>Molecular and Cellular Biology >Mutational analysis of simian virus 40 T antigen: isolation and characterization of mutants with deletions in the T-antigen gene.
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Mutational analysis of simian virus 40 T antigen: isolation and characterization of mutants with deletions in the T-antigen gene.

机译：猿猴病毒40 T抗原的突变分析：T抗原基因缺失的突变体的分离和鉴定。

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摘要

A series of mutants of simian virus 40 has been constructed with deletions in the coding sequence for large T antigen. Nucleotide sequence analysis indicates that 4 mutants have in-phase and 11 have out-of-phase deletions. Mutant DNAs were assayed for the following activities: the ability to form plaques, the ability to produce T antigen as scored by indirect immunofluorescence, viral DNA replication, and morphological transformation of rat cells. Two viable mutants were found, and these had deletions confined to the carboxyl terminus of T antigen. Only those mutants coding for polypeptides greater than 40% of the length of wildtype T antigen produced detectable nuclear fluorescence. The two viable mutants with deletions in the carboxyl terminus of the protein retained the ability both to replicate their DNA, although at a reduced level, and to transform nonpermissive cells. Mutants with sequence changes that result in the loss of more than 117 amino acids from the carboxyl terminus were not viable and were also defective in the DNA replication and transformation functions of T antigen, although several produced detectable nuclear fluorescence. These functions were also sensitive to the removal of amino acids near the amino terminus and in the middle of the protein.
机译：已经构建了一系列猿猴病毒40的突变体，其中大T抗原的编码序列缺失。核苷酸序列分析表明4个突变体具有同相缺失，而11个突变体具有异相缺失。测定突变DNA的下列活性：通过间接免疫荧光，病毒DNA复制和大鼠细胞的形态转化所评分的形成噬斑的能力，产生T抗原的能力。发现了两个可行的突变体，并且这些突变体具有限于T抗原的羧基末端的缺失。仅那些编码大于野生型T抗原长度的40％的多肽的突变体产生可检测的核荧光。蛋白质羧基末端缺失的两个存活突变体既保留了复制其DNA的能力（尽管水平降低了），又保留了转化非许可细胞的能力。尽管有几个产生了可检测的核荧光，但具有序列变化的突变体导致不能从羧基末端丢失超过117个氨基酸，并且它们在T抗原的DNA复制和转化功能方面也存在缺陷。这些功能也对去除氨基酸末端附近和蛋白质中间的氨基酸很敏感。

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