Muller W J, Naujokas M A, Hassell J A
Mol Cell Biol. 1984 Nov;4(11):2406-12. doi: 10.1128/mcb.4.11.2406-2412.1984.
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.
能够产生病毒肿瘤抗原的转化的、允许性小鼠细胞系的存在，将极大地有助于多瘤病毒载体的构建、突变病毒的分析以及整合的多瘤病毒基因组的拯救。为了分离这样的细胞系，我们构建了一个杂交转录单位，其由与多瘤病毒肿瘤抗原编码区融合的猿猴病毒40早期启动子组成。这个杂交转录单位被用于转化NIH 3T3细胞。分离出转化细胞的独立集落，进行再克隆并进行特性分析。在最初分析的10个细胞系中，7个支持携带病毒复制起点的质粒DNA的复制。对其中3个细胞系进行了更详细的特性分析。每个细胞系含有一到两个多瘤病毒DNA的独立插入片段，并合成所有三种病毒肿瘤抗原。此外，三个细胞系中的两个细胞系中的大肿瘤抗原与多瘤病毒复制起点及早期启动子周围的序列特异性结合。这些细胞系不仅对于研究多瘤病毒的复制，而且对于在小鼠细胞环境中研究外源基因的表达都将证明是有用的。
