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Gene Synthesis> | ... Scottsdale, AZ). 2.2. Expression of FLAG-TM3. The gene encoding the EGF domain 4-6 of human thrombomodulin with a FLAG tag (FLAG-TM3) and a C-terminal LPETG motif was designed and synthesized (Genscript Inc.). In ... | Get A Quote |
Introducing unique functional group into protein is an attractive approach for site-selective protein modification applications. In this report, we systemically investigated four site-selective strategies to introduce azide functionality into recombinant thrombomodulin (TM456), via direct recombinant expression with unnatural amino acid, chemical, and enzymatic modification for its bio-orthogonal modification application. First, a straightforward recombinant method to express TM456 with azide functionality near C-terminus by replacing methionine with azidohomoanlanine from methionine auxotroph Escherichia coli cell was investigated. Next, a sortase-mediated ligation (SML) method to incorporate azide functionali... More