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艰难梭菌(p) pppp 合成酶的纯化及体外活性测定

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Immunology and Infection

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JoVE Journal Immunology and Infection

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile

Please note that all translations are automatically generated. Click here for the English version.

艰难梭菌(p) pppp 合成酶的纯化及体外活性测定

DOI:

10.3791/58547-v

•

09:53 min

•

November 03, 2018

•

Astha Pokhrel, Asia Poudel, Erin B. Purcell

¹Department of Chemistry and Biochemistry,Old Dominion University

Chapters

00:04Title
00:44Protein Purification by Nickel Affinity Chromatography
03:30Protein Activity Assay by Thin Layer Chromatography
07:30Results: Thin Layer Chromatography Reveals that C. difficile RSH is an Active ppGpp Synthetase
09:01Conclusion

Summary

Automatic Translation

本文介绍了一种纯化带组氨酸标记的热释蛋白酶的方法, 并利用放射性标记底物和产物的薄层色谱法测定了体外的酶活性。酶活性测定广泛适用于任何激酶、核苷酸循环酶或磷转移反应, 其机理包括三磷酸核苷酸水解。

Tags

Keywords: Protein Purification In Vitro Activity Assay (p)ppGpp Synthetase Clostridium Difficile Phosphotransfer Enzymes Enzyme Substrate Specificity Reaction Kinetics TLC SDS-PAGE Coomassie Blue Staining

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