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Parasitic nematodes cause severely debilitating diseases such as lymphatic filariasis and onchocerciasis. One potential therapeutic target for the treatment or prevention of nematode infections is chitin deacetylase (CDA). CDA converts the biologically important carbohydrate polymer of chitin (beta-1,4-linked N-acetylglucosamine) to chitosan. The effectiveness of this process has been correlated ... read moreto healthy nematode development. The aim of this study was to generate soluble, purified, and active CDA for further characterization through various activity assays. Variations on a bacterial expression system were constructed using the pET22b and pGEX-4T-2 bacterial vectors and the Caenorhabditis elegans CDA gene F48E3.8c. Host cell cultures carrying distinct versions of the recombinant plasmids were extracted using a variety of induction and lysis conditions. Expression of soluble CDA was accomplished using the full F48E3.8c gene, as well as a truncated version of the gene. Enzymatic activity was successfully restored following the denaturation and crude purification of CDA embedded in polyacrylamide gels. Further experiments are required to optimize the purification of active protein in solution.read less
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