Abstract：
To measure the kinetic properties of photorespiratory enzymes, it is necessary to work with purified proteins. Protocols to purify photorespiratory enzymes from leaves of various plant species require several time-consuming steps. It is now possible to produce large quantities of recombinant proteins in bacterial cells. They can be rapidly purified as histidine-tagged recombinant proteins by immobilized metal affinity chromatography using Ni2+-NTA-agarose. This chapter describes protocols to purify several Arabidopsis thaliana His-tagged recombinant photorespiratory enzymes (phosphoglycolate phosphatase, glycolate oxidase, and hydroxypyruvate reductase) from Escherichia coli cell cultures using two bacterial strain-plasmid systems: BL21(DE3)-pET and LMG194-pBAD.
摘要：
为了测量光呼吸酶的动力学特性，有必要使用纯化的蛋白质。从各种植物物种的叶子中纯化光呼吸酶的方案需要几个耗时的步骤。现在可以在细菌细胞中产生大量的重组蛋白。它们可以通过使用Ni2-NTA-琼脂糖的固定化金属亲和层析快速纯化为组氨酸标记的重组蛋白。本章介绍了纯化几种拟南芥His标记的重组光呼吸酶（磷酸乙酸磷酸酶，乙醇酸氧化酶,和羟基丙酮酸还原酶）使用两种细菌菌株-质粒系统从大肠杆菌细胞培养物中获得：BL21（DE3）-pET和LMG194-pBAD。
