Recombinant DNA Technology

1-2-1 units · Undergraduate

Students explore the biochemical basis for the efficacy of a blockbuster drug: Gleevec, which is used to treat chronic myelogenous leukemia. Its target, Abl kinase, is produced in E. coli by recombinant DNA technology, purified using affinity chromatography, and analyzed with polyacrylamide gel electrophoresis, UV-vis spectroscopy, and a colorimetric assay. Natural mutations found in Gleevec-resistant cancer patients are introduced into the ABL1 proto-oncogene with PCR-based mutagenesis and analyzed by agarose gel electrophoresis.