Construction of novel recombinant receptor proteins to improve disease resistance in wheat

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University of the Free State

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English: The aim of the current study was to construct novel recombinant RLK genes encoding chimeric receptor proteins to broaden disease resistance in wheat. Eight different RLKs were initially chosen as extracellular domain donors based on their ability to bind PAMPs/MAMPs or DAMPs. The kinase domain donor was LRK I0, an R protein that renders wheat resistant against Puccinia triticina. These recombinant genes were first designed in silico using a bio-informatic approach where the gene portions encoding the different extracellular domains were linked to the LRKIO kinase domain encoding gene fragment. Translation of the different recombinant genes confirmed that full length polypeptides will be formed. General PAMP binding capacity was therefore combined with the highly specific defence signalling ability of LRKIO within a single protein. Due to several problems encountered, only four recombinant genes were successfully produced, namely BRll-LRKlO, CERK-LRKIO, LecRKI-LRKIO and PERKI-LRKIO. To confirm the success of the approach, LecRKI-RLKIO was successfully transcribed and translated in vitro, thus producing the correct sized polypeptide. Future work will include the production of transgenie wheat to test the ability of these chimeric proteins to increase wheat resistance against pathogen infection.

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