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DpnI (10 U/µL)

DpnI(10U /μL)

Company: Thermo Fisher Scientific
Catalog#: ER1701
Other protocol()

Purification and Crystallization of Chloromuconolactone Dehalogenase ClcF from Rhodococcus opacus 1CP
[Abstract]  The protocol describes the generation of variants of chloromuconolactone dehalogenase from Rhodococcus opacus (R. opacus) 1CP. ClcF is a multimeric protein, which catalyses the dechlorination of 5-chloromuconolactone to cis-dienelactone in the 3-chlorocatecholic acid degradation pathway. The protocol describes the workflow for the purification and subsequent crystallization of the enzyme. The used workflow and the described techniques could be easily adapted to any other protein/enzyme intended to be crystallized by the potential user for subsequent structure determination. The protocol does not involve expensive specialized equipment which allows the use in standard laboratories not specially dedicated to macromolecular crystallography. [摘要]  该方案描述了来自不透明红球菌(emocus)( R。opacus )1CP的氯木聚糖内酯脱卤酶变体的产生。 ClcF是多聚体蛋白,其催化5-氯代粘康酸内酯在3-氯代儿茶酸降解途径中脱氯成顺式 - 二内酯。 该方案描述了酶的纯化和随后的结晶的工作流程。 所使用的工作流程和所描述的技术可以容易地适用于潜在用户为了随后的结构确定而被结晶的任何其它蛋白质/酶。 该协议不涉及昂贵的专用设备,其允许在不专门用于大分子晶体学的标准实验室中使用。