Products/Services Used | Details | Operation |
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Gene Synthesis | … While we originally used point mutations to mutate individual residues one at a time, we would recommend to have the gene synthesized and cloned by a commercial company such as GenScript.com (https://www.genscript.com) or a gBlock (gene block) synthesized by … | Get A Quote |
Intramolecular isopeptide bonds, formed autocatalytically between Lys and Asn/Asp side chains, are widely present in the immunoglobulin-like domains of Gram-positive bacterial adhesins, including Group A Streptococcus, and confer considerable mechanical and chemical stability. These properties make them attractive for applications in biotechnology. Here, we detail the practical considerations that are involved in engineering isopeptide bonds into Ig-like proteins, including the choice of a site where bond-forming residues could be introduced and the appropriate methodology for mutagenesis. We specify how to determine whether an isopeptide bond has formed, what strategies can be adopted to overcome problems, and... More