Uniform 15N-labeled Protein Expression in BL21 ΔE3 E. coli
5:24
Purification of Recombinant Protein from E. coli
7:50
Chemical Attachment of Glucose-5-MTS to Protein by Dialysis
8:48
Results: Site-selective Cys Mutation Followed by In Vitro Glycosylation in STIM1 EF-SAM Protein
10:11
Conclusion
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The overall goal of this procedure is to incorporate cysteine residues in proteins by site directed mutagenesis at natively glycosylated sites and employ in-vitro glucose attachment and assessment approaches to evaluate the efficiency and structur
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Biochemical and structural analyses of glycosylated proteins require relatively large amounts of homogeneous samples. Here, we present an efficient chemical method for site-specific glycosylation of recombinant proteins purified from bacteria by targeting reactive Cys thiols.