U-3'-BCIP: a chromogenic substrate for the detection of RNase A in recombinant DNA expression systems

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...

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Veröffentlicht in:Nucleic acids research 1991-01, Vol.19 (1), p.1-4
Hauptverfasser: Witmer, M R, Falcomer, C M, Weiner, M P, Kay, M S, Begley, T P, Ganem, B, Scheraga, H A
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Sprache:eng
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Zusammenfassung: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/19.1.1