Nishimori K, Kohda T, Segawa K, Oishi M
Institute of Applied Microbiology, University of Tokyo, Japan.
Plasmid. 1991 Nov;26(3):201-8. doi: 10.1016/0147-619x(91)90043-v.
We reported previously that composite DNA constructed from a mammalian plasmid (L factor) and foreign gene can be reestablished as a plasmid in mouse embryonal carcinoma (F9) cells after transfection and the plasmid-bearing F9 cells undergo normal in vitro differentiation in response to retinoic acid, an inducer for F9 cell differentiation. We constructed F9 cells bearing plasmidal L factor DNA in which a reporter (chloramphenicol acetyltransferase; CAT) gene was placed under the control of a differentiation-responsive viral (Moloney murine leukemia virus or simian virus 40) enhancer-promoter. When such plasmid-bearing cells were treated with retinoic acid, the CAT gene was inducibly expressed. These results indicate that mammalian gene expression can be studied with the plasmidal expression vector which is structurally dissociated from complex chromosomes.
我们先前报道,由哺乳动物质粒(L因子)和外源基因构建的复合DNA在转染后可在小鼠胚胎癌细胞(F9)中重新建立为质粒,并且携带该质粒的F9细胞在视黄酸(F9细胞分化诱导剂)作用下经历正常的体外分化。我们构建了携带质粒L因子DNA的F9细胞,其中报告基因(氯霉素乙酰转移酶;CAT)置于分化反应性病毒(莫洛尼鼠白血病病毒或猿猴病毒40)增强子-启动子的控制之下。当用视黄酸处理这些携带质粒的细胞时,CAT基因可被诱导表达。这些结果表明,可利用与复杂染色体结构分离的质粒表达载体来研究哺乳动物基因表达。