Generation of a tissue transglutaminase expression library by C-Terminal nested deletion Custom Essay

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Tissue transglutaminase (TG2) is a protein cross-linking enzyme that stabilises structures in the body such as blood clots, hair, skin and the extracellular matrix. It has been implicated in many different diseases and is a potential therapeutic target. As the antigen in celiac disease, there are a large number of TG2-reactive antibodies that have been identified by antibody phage display. However, most of these have not had their TG2 epitopes mapped. This project will generate a library of C-terminally truncated TG2 mutants, each with a different sized deletion. Expression of these mutants in E. coli will facilitate the rapid identification of TG2 epitopes and the characterisation of celiac disease antibodies.
Will use molecular biology techniques, including nested deletion using the ExoIII/SI method, PCR, cloning. Also affinity purification, biochemical assays, SDS-PAGE, Western blotting

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