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|Title:||Preparation of functional human urokinase proteins|
|Abstract:||Human urokinase is produced using recombinant DNA techniques. The invention disclosed thus enables the production of urokinase free of contaminants with which it is ordinarily associated in its native cellular environment. Methods, expression vehicles and various host cells useful in its production are also disclosed.|
|Inventor(s):||Heyneker; Herbert L. (Burlingame, CA), Holmes; William E. (San Mateo, CA), Vehar; Gordon A. (San Carlos, CA)|
|Assignee:||Genentech, Inc. (South San Francisco, CA)|
|Filing Date:||Jul 19, 1988|
|Claims:||1. A DNA isolate consisting essentially of a DNA sequence encoding a human urokinase. |
2. The DNA sequence according to claim 1 operably linked with a DNA sequence capable of effecting expression thereof.
3. A recombinant expression vector containing a DNA sequence encoding a human urokinase wherein the vector is capable, in a transformant microorganism or cell culture, of expressing said DNA sequence.
4. The vector according to claim 3, said human urokinase having about 411 amino acids as set forth in FIG. 4.
5. The vector according to claim 3, said human urokinase having about 279 amino acids as set forth in FIG. 2.
6. The DNA vector according to claim 3, wherein the DNA sequence encodes the amino acid sequence Lys-Ile-Ile-Gly-Gly at codons 158-162 taking codon 1 as that encoding the first amino acid of the mature form of human urokinase.
7. A microorganism transformed with the vector according to claim 3, said microorganism being capable of producing by expression a human urokinase.
8. A microorganism according to claim 7 obtained by transforming an E. coli strain.
9. The microorganism according to claim 7, said human urokinase having about 411 amino acids as set forth in FIG. 4.
10. A cell culture capable of expressing a human urokinase, obtained by transforming a mammalian cell line with a vector according to claim 3.
11. The cell culture according to claim 10, said human urokinase having about 411 amino acids as set forth in FIG. 4.
12. A process which comprises expressing a DNA sequence encoding a human urokinase in a recombinant host cell, said recombinant host cell obtained by transforming a microorganism or cell culture with an expression vector containing said DNA sequence.
13. A process which comprises growing in a growth medium recombinant cells obtained by transforming a microorganism or cell culture with an expression vector containing DNA encoding a human urokinase and simultaneously expressing said DNA.
14. A process comprising transforming a microorganism or cell culture with a replicable vector containing recombinant DNA encoding a human urokinase and expressing said DNA.
15. A process according to any one of claims 12, 13 or 14 wherein said microorganism is E. coli.
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