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. 2004 Aug 31;101(37):13454–13459. doi: 10.1073/pnas.0405521101

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Copyright © 2004, The National Academy of Sciences

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Fig. 5. — tmRNA-mediated tagging. (a) Cartoon of tagging of the λ-cI-N-trpAt protein. Leadered λ-cI-N-trpAt mRNA is translated in an S1-dependent manner, whereas the leaderless mRNA is translated in an S1-independent manner. Translation of both types of mRNA leads to a stalled ribosome complex, which is resolved by tmRNA-mediated tagging or ribosomal fall-off, resulting in tagged and untagged λ-cI-N-trpAt protein. (b) Coomassie-stained SDS gel of λ-cI-N-trpAt proteins isolated by nickel affinity after a tagging assay with leadered λ-cI-N-trpAt mRNA in which S1 variants were overexpressed. Tagged and untagged proteins are indicated by arrows. (c) λ-cI-N-trpAt proteins from tagging experiments employing leaderless and extended SD λ-cI-N-trpAt mRNA.