Figure 3
From: Enabling stop codon read-through translation in bacteria as a probe for amyloid aggregation
At least two WH1(Rn) repeats fused to RF1 are required to enable stop codon read-through. (a) Serial dilutions of bacterial cultures on LB-agar expressing the reporters lacZ-WT (top) or lacZ-amber (bottom), a variant carrying a premature stop codon, in the presence of a chromogenic substrate (X-Gal). Aggregation of the WH1(R2-4)-RF1 chimeras at 42 °C promotes amber codon read-through, while the chimeras that remain soluble, WH1(R0-1)-RF1, efficiently promote the end of translation. (b) A liquid culture assay in supplemented minimal medium. Again, blue colour (stop codon read-through) is observed for the WH1(R2-4)-RF1 chimeras in the lacZ-amber background. (c) β-galactosidase activity was determined in cells expressing the WH1(R0-4)-RF1 chimeras. Values show the percentage of activity for each chimera in combination with the lacZ-amber reporter, relative to the lacZ-WT reporter (inset). The histograms display the media and SDs (whiskers) from 4 independent replicas.
