U-3'-BCIP：一种用于在重组DNA表达系统中检测核糖核酸酶A的显色底物。
U-3'-BCIP: a chromogenic substrate for the detection of RNase A in recombinant DNA expression systems.
作者信息
Witmer M R, Falcomer C M, Weiner M P, Kay M S, Begley T P, Ganem B, Scheraga H A
机构信息
Baker Laboratory of Chemistry, Cornell University, Ithaca, NY 14853-1301.
出版信息
Nucleic Acids Res. 1991 Jan 11;19(1):1-4. doi: 10.1093/nar/19.1.1.
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.
摘要
本文描述了牛胰核糖核酸酶A（RNase A，EC 3.1.27.5）的显色底物尿苷-3'-（5-溴-4-氯吲哚-3-基）-磷酸酯（U-3'-BCIP）的合成。RNase A催化U-3'-BCIP水解，释放出卤代吲哚-3-醇，该醇迅速进行有氧氧化生成深蓝色的5,5'-二溴-4,4'-二氯靛蓝。初步动力学研究表明，该化合物可能在体外和体内测定RNase A活性方面具有实际用途，例如在筛选通过重组DNA方法产生RNase A的细菌菌落时。
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