TY - JOUR
T1 - Amplification of P450c21 expression in cultured mammalian cells
AU - Ricketts, Michael H.
AU - Chiao, Eric
AU - Hu, Meng Chun
AU - Chung, Bon chu
N1 - Funding Information:
We thank Dr. Lih-Hwa Hwang (Development Center for Biotechnology, Taiwan) and Genentech Inc. (California, U.S.A.) for the gifts of pCMV-3, Rat-1 cells and pFD11. This work was supported by National Science Council (NSC #79-0412-B001-16 and #NSC-79-0412-B001-19) and Academia Sinica, Republic of China.
PY - 1992/7/15
Y1 - 1992/7/15
N2 - 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.
AB - 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.
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U2 - 10.1016/S0006-291X(05)80825-3
DO - 10.1016/S0006-291X(05)80825-3
M3 - Article
C2 - 1321611
AN - SCOPUS:0026768246
SN - 0006-291X
VL - 186
SP - 426
EP - 431
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -