| CPC G16B 15/30 (2019.02) [C12N 9/1205 (2013.01)] | 17 Claims |
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1. A method for selection of a target plant lysin motif (LysM) receptor and modifying the target plant LysM receptor to have a desired receptor characteristic, comprising:
(a) providing a structural model, a molecular model, a surface characteristics model, an electrostatic potential model, or combinations thereof of a donor plant LysM receptor having the desired receptor characteristic and two or more potential target plant LysM receptors, wherein the desired receptor characteristic is affinity, selectivity, specificity, or combinations thereof for an oligosaccharide or class of oligosaccharides;
(b) comparing each of the two or more potential target plant LysM receptors with the structural model, the molecular model, the surface characteristics model, the electrostatic potential model of the donor plant LysM receptor, or combinations thereof; or comparing each of the two or more potential target plant LysM receptors with the donor plant LysM receptor using structural overlay;
(c) selecting the potential target plant LysM receptor for modification;
(d) identifying one or more amino acid residues for modification in the target LysM receptor by comparing amino acid residues of a first oligosaccharide binding feature in the donor plant LysM receptor with the corresponding amino acid residues in the target plant LysM receptor; and
(e) generating a modified plant LysM receptor wherein the one or more amino acid residues in the first oligosaccharide binding feature of the target plant LysM receptor have been substituted with corresponding amino acid residues from the donor plant LysM receptor; wherein (i) the one or more amino acid residues in the first oligosaccharide binding feature include amino acids corresponding to SEQ ID NO: 63 and SEQ ID NO: 64 of Lotus japonicus NFR5, (ii) the one or more amino acid residues in the first oligosaccharide binding feature include amino acids corresponding to L147, L151, L152, L154, T156, K157, and V158 of Medicago NFP (SEQ ID NO: 1), (iii) the one or more amino acid residues in the first oligosaccharide binding feature include amino acids corresponding to SEQ ID NO: 28 and SEQ ID NO: 29 of Lotus NFR1 and/or SEQ ID NO: 30 and SEQ ID NO: 31 of Lotus CERK6, or (iv) the one or more amino acid residues in the first oligosaccharide binding feature include combinations of (i)-(iii).
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