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. 2002 May 25;288(3):305–320. doi: 10.1006/jmbi.1999.2688

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Copyright © 1999 Academic Press. All rights reserved.

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Influence of stem 1 length on −1 ribosomal frameshifting (part II). (a) The stem 1 base composition of members of the pSL series. Mutations were introduced into stem 1 of constructs pFScass 5, 6 or 7 such that stem 1 length was increased or decreased in single base-pair steps. The predicted stability of stem 1 of each construct is indicated. Also shown is the size of the −1 frameshift product specified by each frameshift signal and the efficiency of frameshifting seen in RRL. All constructs possess the same loop 1, loop 2 and stem 2 sequences as pFScass 6 except plasmid pSL8, a variant of pSL7 with a destabilised stem 2. (b) RRL translation products synthesised in response to mRNAs derived from BamHI-digested pSL1 and mutant derivatives. Products were labelled with [³⁵S]methionine, separated on a SDS-15 % polyacrylamide gel and detected by autoradiography. The frameshifted (22, 28 or 85 kDa) and non-frameshifted (19 kDa) species are marked with arrows. For the pSL series, mRNAs were translated at two concentrations (25 μg/ml and 12.5 μg/ml). Track M was loaded with [¹⁴C] high molecular mass markers (Amersham) and a control translation programmed with water was also included.

Influence of stem 1 length on −1 ribosomal frameshifting (part II). (a) The stem 1 base composition of members of the pSL series. Mutations were introduced into stem 1 of constructs pFScass 5, 6 or 7 such that stem 1 length was increased or decreased in single base-pair steps. The predicted stability of stem 1 of each construct is indicated. Also shown is the size of the −1 frameshift product specified by each frameshift signal and the efficiency of frameshifting seen in RRL. All constructs possess the same loop 1, loop 2 and stem 2 sequences as pFScass 6 except plasmid pSL8, a variant of pSL7 with a destabilised stem 2. (b) RRL translation products synthesised in response to mRNAs derived from BamHI-digested pSL1 and mutant derivatives. Products were labelled with [³⁵S]methionine, separated on a SDS-15 % polyacrylamide gel and detected by autoradiography. The frameshifted (22, 28 or 85 kDa) and non-frameshifted (19 kDa) species are marked with arrows. For the pSL series, mRNAs were translated at two concentrations (25 μg/ml and 12.5 μg/ml). Track M was loaded with [¹⁴C] high molecular mass markers (Amersham) and a control translation programmed with water was also included.