Katharine Schalk Kramer

US Patent:

61366056, Oct 24, 2000

Filed:

Aug 26, 1994

Appl. No.:

8/297431

Inventors:

William E. Fahl - Madison WI
Andrew M. Gulick - Madison WI
T. Herbert Manoharan - Madison WI
Ralph B. Puchalski - La Jolla CA
Katharine Kramer - Madison WI
Wyeth W. Wasserman - Madison WI

Assignee:

Wisconsin Alumni Research Foundation - Madison WI

International Classification:

C12N 1500
C12N 1512
C07H 2104

US Classification:

435440

Abstract:

A method is described for the creation of novel isoforms of the enzyme glutathione S-transferase which have enhanced activity in host cells against specific toxic agents. The method includes site directed mutagenesis and selection with the targeted agent in the host cells. The sites of directed mutagenesis is the site of electrophile binding by the native form of the enzyme. This site has proven susceptible to manipulation without loss of enzymatic activity. Various techniques for enhancing the expression, activity, or localization of the expressed enzyme in mammalian cells are described. Genes for the mutant isoforms of the enzyme may be useful in cancer therapeutics to confer upon selected groups of cells heightened resistance to antineoplastic agents.