Hi everyone I'm interested in making a synthetic pathogen sensor using human toll- like receptors 2 and 6 The idea is to ligate the pathogen-sensing, extracellular domains of each TLR (or possibly just the recognition module, consisting of just a single residue) to one half of a GFP, producing a visible signal when the TLRs dimerise. Using a prokaryotic chassis to carry the TLRs might not be possible owing to the absence of endoplasmic reticula in prokaryotes, and the consequent problems with protein folding, as well as with being able to anchor the transmembrane domains of the TLRs in the cell wall. Any TLRs expressed might not fold properly. I think that TLR signal transduction is not well understood hence the use of GFPs as reporters, as bacteria would lack the requisite intracellular signalling pathways. If these problems prove insuperable, then a eukaryotic chassis might have to be used. Anyway, it's a learning curve!
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