PCR Mutagenesis, Cloning, Expression, Fast Protein Purification Protocols and Crystallization of the Wild Type and Mutant Forms of Tryptophan SynthaseEduardo Hilario 1, Li Fan 2, Leonard J. Mueller 1, Michael F. Dunn 2
1Department of Chemistry, University of California-Riverside, 2Department of Biochemistry, University of California-Riverside
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.