Method for generating proteolytic enzymes specific against a sel

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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435220, C12N 950, C12N 952

Patent

active

056020210

ABSTRACT:
The present invention describes methods for screening or selecting novel proteolytic enzymes. These enzymatic functions can be selected from isolated chromosomal DNA libraries or from pools of mutagenized DNA encoding a proteolytic function. In particular, the selection method of the present invention, a phage gene is chosen that encodes a gene product necessary for the production of a phage. The phage carrying the modified gene is introduced into a host. Also, DNA libraries present within a cloning vector are introduced into host cells. The host cells are grown under conditions where the introduced libraries are expressed in the host cells. The presence of an enzymatic function capable of cleaving the target peptide is identified on the basis of production of infective phage.

REFERENCES:
patent: 5258289 (1993-11-01), Davis et al.
Matthew, D. J., and J. A. Wells, "Substrate Phage: Selection of Protease Substrates by Monovalent Phage Display," Science 260: 1113-1117 (1993).
Roberts, B. L., et al., "Protease inhibitor display M13 phage: selection of high-affinity neutrophil elastase inhibitors," Gene 121: 9-15 (1992).

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