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Overexpression and Purification of Human Cis-prenyltransferase in Escherichia coli
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1Sackler Faculty of Medicine,Tel Aviv University, 2Department of Physiology and Pharmacology, Sackler Faculty of Medicine,Tel Aviv University, 3Department of Ophthalmology, Tel Aviv Sourasky Medical Center, affiliated to the Sackler Faculty of Medicine,Tel Aviv University, 4Tel Aviv Sourasky Medical Center, affiliated to the Sackler Faculty of Medicine,Tel Aviv University
Chapters
- 00:05Title
- 00:56Purification of Human DHDDS
- 03:52Analytical Size-exclusion Chromatography (SEC)
- 04:40Enzyme Kinetics – Time-dependent Activity
- 06:15Results: Purification and Analysis of Human DHDDS
- 07:00Conclusion
A simple protocol for overexpression and purification of codon-optimized, human cis-prenyltransferase, under non-denaturing conditions, from Escherichia coli, is described, along with an enzymatic activity assay. This protocol can be generalized for production of other cis- prenyltransferase proteins in quantity and quality suitable for mechanistic studies.
