Human t-PA production using vectors coding DHFR protein

Details Human t-PA production using vectors coding DHFR protein Human t-PA production using vectors coding DHFR protein

1990-03-22

1991-04-23

Martinell, James

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 91, 4351723, 4352402, 4353201, C12N 1500, C12N 516, C12P 2102

Patent

active

050100025

ABSTRACT:
A method for producing tissue plasminogen activator (t-PA) in eukaryotic host cells is disclosed. Enhanced levels of t-PA production are obtained by co-amplification of the t-PA gene through treatment of cultures transformed with mutant or wild type DHFR with methotrexate.

REFERENCES:
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patent: 4419446 (1983-12-01), Howley et al.
Odenakker et al., "Messenger RNA for Human Tissue Plasminogen Activator", Eur. J. Biochem. 121: 269 (1982).
Haber et al., "Unstable Amplification of an Altered Dihydrofolate Reductase Gene Associated with Double-Minute Chromosomes", Cell 26: 355 (1981).
Chang et al., "Phenotypic Expression in E. coli of a DNA Sequence Coding for Mouse Dihydrofolate Reductase", Nature 275: 617 (1978).
Blattner et al.; "Cloning Human Fetal Gamma-Globin and Mouse Alpha-Type Globin DNA: Preparation and Screening of Shotgun Collections", Science 202: 334 (1978).
Christman, Judith K. et al., Proc. Natl. Acad. Sci. 79: 1815 (1982).
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Ringold, Gordon et al., Journal of Molecular and Applied Genetics 1: 165 (1981).
Wigler et al., Proc. Natl. Acad. Sci. (U.S.A.) 77, 3567 (1980).

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