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Mol Cell Biol. 1982 September; 2(9): 1044-1051
Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
C M Gorman,
L F Moffat and
B H Howard
ABSTRACT
We constructed a series of recombinant genomes which directed expression of the enzyme chloramphenicol acetyltransferase (CAT) in mammalian cells. The prototype recombinant in this series, pSV2-cat, consisted of the beta-lactamase gene and origin of replication from pBR322 coupled to a simian virus 40 (SV40) early transcription region into which CAT coding sequences were inserted. Readily measured levels of CAT accumulated within 48 h after the introduction of pSV2-cat DNA into African green monkey kidney CV-1 cells. Because endogenous CAT activity is not present in CV-1 or other mammalian cells, and because rapid, sensitive assays for CAT activity are available, these recombinants provided a uniquely convenient system for monitoring the expression of foreign DNAs in tissue culture cells. To demonstrate the usefulness of this system, we constructed derivatives of pSV2-cat from which part or all of the SV40 promoter region was removed. Deletion of one copy of the 72-base-pair repeat sequence in the SV40 promoter caused no significant decrease in CAT synthesis in monkey kidney CV-1 cells; however, an additional deletion of 50 base pairs from the second copy of the repeats reduced CAT synthesis to 11% of its level in the wild type. We also constructed a recombinant, pSV0-cat, in which the entire SV40 promoter region was removed and a unique HindIII site was substituted for the insertion of other promoter sequences.
Mol Cell Biol. 1982 September; 2(9): 1044-1051
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Cha, J.-y., Kim, H.-s., Kim, H.-i., Im, S.-s., Kim, S.-y., Kim, J.-w., Yeh, B.-i., Ahn, Y.-h.
(2002). Analysis of Polymorphism of the GLUT2 Promoter in NIDDM Patients and its Functional Consequence to the Promoter Activity. Annals of Clinical & Laboratory Science
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Akiyama, T. E., Baumann, C. T., Sakai, S., Hager, G. L., Gonzalez, F. J.
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Rohrdanz, E., Ohler, S., Tran-Thi, Q.-H., Kahl, R.
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Endo, K., Hirata, A., Iwai, K., Sakurai, M., Fukushi, M., Oie, M., Higuchi, M., Hall, W. W., Gejyo, F., Fujii, M.
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Garcia, B., Santisteban, P.
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Drover, V. A. B., Wong, N. C. W., Agellon, L. B.
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Medina, D. L., Rivas, M., Cruz, P., Barroso, I., Regadera, J., Santisteban, P.
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Lopez, N., Jacamo, R., Franze-Fernandez, M. T.
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Liu, X., Wall, Q.-T., Taylor, L., Curthoys, N. P.
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Piaggio, E., Sanceau, J., Revelli, S., Bottasso, O., Wietzerbin, J., Serra, E.
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