Amplification of P450c21 expression in cultured mammalian cells.
We describe in this paper an investigation of mammalian expression systems for P450c21 (21-hydroxylase). Four different promoters, the SV40 early and late promoters, MMTV-LTR, and CMV immediate early promoter were tested for their ability to drive the expression of P450c21 in cultured COS-1 cells. With the exception of MMTV-LTR, all drove the expression of similar levels of functional 21-hydroxylase. In addition, the Rat-1 cell line was tested and shown to be suitable for the stable expression of functional P450c21. We have established cell lines derived from Rat-1 either normal or mutant P450c21 stably expressed together with amplifiable markers. The expression of P450c21 was further increased by selection in methotrexate.
Ricketts
M H
MH
Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, Taiwan, Republic of China.
Chiao
E
E
Hu
M C
MC
Chung
B C
BC
D000818:Animals; D002460:Cell Line; D002999:Clone Cells; D005784:Gene Amplification; D005809:Genes, Regulator; D007700:Kinetics; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics; D013255:Steroid 21-Hydroxylase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
D013255:Steroid 21-Hydroxylase