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PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
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PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan Synthase
Capítulos
- 00:04Introduction
- 00:44Purification of Wild Type or Mutant Form of α2β2 StTS Complex
- 03:53Optimized Crystallization for Wild Type and Mutant Form of the α2β2 StTS Comple
- 06:49Results: Purification of the α- and β -Subunits of the Tryptophan Synthase and the Wild Type and Mutant α2β2 StTS Complexes
- 08:50Conclusion
This article presents a series of consecutive methods for the expression and purification of Salmonella typhimurium tryptophan synthase comp this protocol a rapid system to purify the protein complex in a day. Covered methods are site-directed mutagenesis, protein expression in Escherichia coli, affinity chromatography, gel filtration chromatography, and crystallization.
