PubMed · 10453638
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B Steipe. 1999. Evolutionary approaches to protein engineering.. https://doi.org/10.1007/978-3-642-60142-2_4
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Engineering gene circuits with novel functions holds promise for broad applications in biology, engineering, and medicine. Directed evolution complements rational design as an important strategy for optimizing gene circuits and circuit elements.
A major class of disease-resistance (R) genes in plants encode nucleotide-binding site/leucine-rich repeat (LRR) proteins. The LRR domains mediate recognition of pathogen-derived elicitors. Here we describe a random in vitro mutation analysis illustrating how mutations in an R protein (Rx) LRR domain generate disease-resistance specificity. The original Rx protein confers resistance only against a subset of potato virus X (PVX) strains, whereas selected mutants were effective against an additional strain of PVX and against the distantly related poplar mosaic virus. These effects of LRR mutations indicate that in vitro evolution of R genes could be exploited for enhancement of disease resistance in crop plants. Our results also illustrate how short-term evolution of disease resistance in wild populations might be toward broader spectrum resistance against multiple strains of the pathogen. The breadth of the disease-resistance phenotype from a natural R gene may be influenced by the tradeoff between the costs and benefits of broad-spectrum disease resistance.
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