Bioautographic visualization of dihydrofolate reductase in enzyme overproducing BHK mutants |
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| Authors: | S L Naylor J K Townsend R J Klebe |
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| Institution: | (1) Division of Human Genetics, Department of Human Biological Chemistry and Genetics, The University of Texas Medical Branch, 77550 Galveston, Texas;(2) Present address: Biochemical Genetics Section, Roswell Park Memorial Institute, 14263 Buffalo, New York |
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| Abstract: | A bioautographic procedure has been developed for the visualization of the isozymes of dihydrofolate reductase (DHFR, E.C. 1.5.1.3). In addition to detecting electrophoretically separated enzymes, bioautography was utilized to visualize DHFR after isoelectric focusing on polyacrylamide gels. Both zone electrophoresis and isoelectric focusing were used to compare wildtype BHK cells to mutants which overproduce dihydrofolate reductase. In agreement with other physical data, the BHK-A5 overproduction mutant appears to produce more dihydrofolate reductase of the same electrophoretic mobility and isoelectric point as wild type cells.This study was supported by Grants GM 21433 and CA 19019 from the National Institutes of Health. |
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| Keywords: | dihydrofolate reductase mutant cells bioautography electrophoresis |
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