Transcription of Champion pET SUMO Protein Expression System
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User Manual Champion pET SUMO Protein Expression System For high-level Expression and enhanced solubility of recombinant proteins in E. coli and cleavage of native Protein Catalog no. K300-01 Rev. Date: 18 June 2010 Manual part no. 25-0709 MAN0000440 ii iii Table of Contents Kit Contents and Storage ..v Accessory T7-Regulated Expression ..3 Experimental Outline ..5 Methods .. 6 Cloning Considerations ..6 Producing PCR Products ..8 Cloning into pET Transforming One Shot Mach1 -T1R Competent Cells ..10 Analyzing Expressing the PCR Product ..13 Analyzing Samples ..16 Purifying the Recombinant Fusion Using SUMO Troubleshooting.
pET Expression System The Champion™ pET Expression System is based on expression vectors originally developed by Studier and colleagues, and takes advantage of the high activity and specificity of the bacteriophage T7 RNA polymerase to allow regulated expression of heterologous genes in E. coli from the T7 promoter
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