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U-3'-BCIP: a chromogenic substrate for the detection of RNase A in recombinant DNA expression systems
M R Witmer1, C M Falcomer, M P Weiner
1Baker Laboratory of Chemistry, Cornell University, Ithaca, NY 14853-1301.
Nucleic Acids Research
|January 11, 1991
Abstract:
The synthesis of the bovine pancreatic ribonuclease A (RNase A, EC 3.1.27.5) chromogenic substrate uridine-3'-(5-bromo-4-chloroindol-3-yl)-phosphate (U-3'-BCIP) is described. RNase A catalyzes the hydrolysis of U-3'-BCIP to release a halogenated indol-3-ol that undergoes rapid aerobic oxidation to the dark blue 5,5'-dibromo-4,4'-dichloroindigo. Preliminary kinetic studies indicate that this compound may have practical use for assaying RNase A activity both in vitro and in vivo, e.g. in screening bacterial colonies for RNase A produced by recombinant DNA methods.