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. 2009 Dec 3;38(4):1341–1352. doi: 10.1093/nar/gkp1073

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© The Author(s) 2009. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Non-AUG initiation, stop codon read-through and reading frame maintenance in the absence of rsmH and/or rsmI. (A) Efficiency of translation initiation at non-AUG codons in E. coli wild-type (WT) (white), ΔrsmI (gray), ΔrsmH (stripe) and ΔrsmI/ΔrsmH (black). The reporter constructs with various initiation codons are shown at the top. The reporter genes Rluc (Renilla), starting with AUG codon, and Fluc (Firefly), starting with various codons (AUG, AUU, UUG, GUG or AAA), are tandemly arranged in the same transcript. The efficiency of translation initiation was assessed by dividing the chemiluminescence of Fluc by that of Rluc (the F/R value). For each initiation codon, the relative F/R values for the knockout strains were normalized to that of the wild-type strain. SD: Shine–Dalgarno sequence. (B) Assessment of stop codon read-through and reading frame maintenance in E. coli wild-type (WT) (white), ΔrsmI (gray), ΔrsmH (stripe) and ΔrsmI/ΔrsmH (black). The reporter constructs shown at the top contain the Rluc (Renilla) and Fluc (Firefly) open reading frames with short windows to assess stop codon read-through (UGA or UAG) or reading frame maintenance (+1 or –1 frameshifts) inserted between them. The translational efficiency of each construct was measured by the F/R value. For each construct, the relative F/R values for the knockout strains were normalized to that of the wild-type strain.