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首页> 外文期刊>Molecular and Cellular Biology >Isolation of large T antigen-producing mouse cell lines capable of supporting replication of polyomavirus-plasmid recombinants.
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Isolation of large T antigen-producing mouse cell lines capable of supporting replication of polyomavirus-plasmid recombinants.

机译：能够支持多瘤病毒质粒重组体复制的大型T抗原产生小鼠细胞系的分离。

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Construction of polyomavirus vectors, analysis of mutant viruses, and rescue of integrated polyomavirus genomes would be considerably aided by the availability of transformed, permissive mouse cell lines capable of producing the viral tumor antigens. To isolate such cell lines, we constructed a hybrid transcription unit composed of the simian virus 40 early promoter fused to the coding region for the polyomavirus tumor antigens. This hybrid transcription unit was used to transform NIH 3T3 cells. Independent foci of transformed cells were isolated, recloned, and characterized. Among 10 lines initially analyzed, 7 supported the replication of origin-bearing plasmid DNAs. Three cell lines were characterized in greater detail. Each line contained one or two independent insertions of polyomavirus DNA and synthesized all three viral tumor antigens. Moreover, the large tumor antigen in two of three lines bound with specificity to sequences about the polyomavirus origin and early promoter. These cell lines should prove useful for studying not only the replication of polyomavirus but also the expression of foreign genes in a mouse cell environment.
机译：多瘤病毒载体的构建，突变病毒的分析以及整合多瘤病毒基因组的挽救将大大受益于能够产生病毒肿瘤抗原的转化的，允许的小鼠细胞系的可用性。为了分离这样的细胞系，我们构建了由猿猴病毒40个早期启动子组成的杂合转录单位，所述猿猴病毒40个早期启动子与多瘤病毒肿瘤抗原的编码区融合。该杂交转录单位用于转化NIH 3T3细胞。分离，克隆和表征转化细胞的独立灶。在最初分析的10条品系中，有7条支持带有起点的质粒DNA的复制。对三种细胞系进行了更详细的表征。每个品系包含一个或两个独立的多瘤病毒DNA插入片段，并合成了所有三种病毒性肿瘤抗原。而且，三株中的两株中的大肿瘤抗原与多瘤病毒起源和早期启动子的序列特异性结合。这些细胞系不仅对研究多瘤病毒的复制而且对研究小鼠细胞环境中外源基因的表达都将是有用的。

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