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. 2006 Feb 25;34(4):1250–1260. doi: 10.1093/nar/gkl017

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© The Author 2006. Published by Oxford University Press. All rights reserved

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Mutational analysis of a downstream stimulator on frameshifting efficiencies mediated by the hepta- and octo-uridine stretches. (a) Diagram showing the slippery sequence and the downstream stem–loop structures of SARS-CoV 1a/1b region. The slippery sequence is underlined and the two stems are indicated. Also indicated are the mutations introduced into stem 2 (in bold). (b) Expression of pF-S1ab (lane 1), pF-S1abM (lane 2), pF-S1ab/7T (lane 6), pF-S1ab/7TM (lane 7), pF-S1ab/8T (lane 3) and pF-S1ab/8T (lane 4) in Cos-7 cells. Cells were infected with the recombinant vaccinia/T7 virus, and transfected with an empty control plasmid (lane 5) and the six constructs, respectively. At 18 h posttransfection, cells were harvested and lysates prepared. Polypeptides were separated on SDS–12% polyacrylamide and analyzed by western blot with anti-Flag antibodies. Bands corresponding to 1a (or 1ab), the frameshifting products (FS) and β-tubulin are indicated. Numbers on the left indicate molecular masses in kilodaltons.