AZOTOBACTER VINELANDII RNA POLYMERASE .6. AN IN SITU ASSAY FOR POLYMERASE AFTER ACRYLAMIDE GEL ELECTROPHORESIS

Analysis of the A. vinelandii RNA polymerase by acrylamide gel electrophoresis indicated that enzyme activity was associated with more than one of the resolved protein bands. Initially RNA polymerase activity was determined after elution from gels sliced into discs (Chrambach, 1966). However, recovery of enzyme activity was variable and closely spaced protein bands could not be consistently assayed separately. The present paper describes a procedure for assaying for RNA polymerase in the acrylamide gel. This assay utilizes the unprimed synthesis of poly A·poly U (Smith et al, 1967; Krakow, 1968) or the rIC copolymer (Krakow and Karstadt, 1967);followed by staining with ethidium bromide to visualize the polymers. © 1968.