Kaplan D R, Bockus B, Roberts T M, Bolen J, Israel M, Schaffhausen B S
Mol Cell Biol. 1985 Jul;5(7):1795-9. doi: 10.1128/mcb.5.7.1795-1799.1985.
A recombinant plasmid containing a metallothionein promoter-polyoma middle T cDNA fusion was constructed and used to transfect NIH 3T3 cells. Transformed cells expressing middle T were injected into nude mice. Within 3 weeks, each mouse produced tumors containing middle T equivalent to that in 250 to 1,000 100-mm dishes of polyomavirus-infected cells. This middle T, partially purified by immunoaffinity chromatography, retained activity as measured by its ability to be phosphorylated in vitro. The combined approach of fusing strong promoters to genes of interest and utilizing nude mice to grow large quantities of cells expressing the gene provides a quick, inexpensive alternative to other expression systems.
构建了一个含有金属硫蛋白启动子-多瘤病毒中间T cDNA融合体的重组质粒，并用于转染NIH 3T3细胞。将表达中间T的转化细胞注射到裸鼠体内。在3周内，每只小鼠都产生了含有中间T的肿瘤，其含量相当于250至1000个100毫米培养皿中感染多瘤病毒的细胞中的含量。通过免疫亲和层析部分纯化的这种中间T，通过其在体外被磷酸化的能力来衡量，保留了活性。将强启动子与感兴趣的基因融合并利用裸鼠大量培养表达该基因的细胞的联合方法，为其他表达系统提供了一种快速、廉价的替代方法。
