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. 2006 Apr 28;103(19):7518–7523. doi: 10.1073/pnas.0602379103

Fig. 5.

Fig. 5.

Transformants of susceptible barley cv. Golden Promise with mutated domain pK1 or pK2. The two lysines in the ATP anchor were mutated to N and Q. (A) Southern blot of HindIII-digested barley DNA hybridized with the 4.5-kb genomic probe, RSB228. Lane 1, cv. Morex; lanes 2–12, T2 transformants containing a single copy of the mutated pK1 transgene of pK1; lanes 13 and 14, T1 transformants containing three copies of the mutated pK2 transgene of pK2; lane 15, cv. Golden Promise. (B) Western blot demonstrating that all lines except cv. Golden Promise express the Rpg1 protein. (C) Autophosphorylation of in planta synthesized Rpg1. Rpg1 was immunoprecipitated from the same leaf samples scored for disease and subjected to autophosphorylation. Rpg1 from the mutated pK1 transformants exhibited autocatalytic activity (lanes 2–12), whereas Rpg1 of the mutated pK2 transformants lost the ability to autophosphorylate (lanes 13 and 14).