Protein amount of DenA-GFP and amino acid substituted variants during development.

Abstract

<p>Western hybridization with equal amounts of protein extracts of DenA-GFP (54.5 kDa) compared to <b>(A)</b> DenA<sup>S253D</sup>-GFP carrying a negative charge reminiscent of a phosphorylated protein, <b>(B)</b> DenA<sup>S253A</sup>-GFP which cannot be phosphorylated, <b>(C)</b> DenA<sup>S243D-S245D-S253D</sup>-GFP with three negative charges mimicking a triple phosphorylated protein and <b>(D)</b> the corresponding DenA<sup>S243A-S245A-S253A</sup>-GFP which cannot be phosphorylated. Samples were taken from vegetative hyphae (Veg) and at indicated time points (in hours) of illumination which induces asexual development (Asex). Membranes were treated with GFP-antibody to visualize the fusion protein and free GFP (25 kDa). Loading control: Ponceau staining. Lower panels show quantification of band intensities of DenA-GFP and its respective variants relative to vegetative growth.</p

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