In vivo characterisation of a translational enhancer upstream from the coat protein open reading frame of potato virus S.
The 101 nucleotide region upstream from the ATG of the potato virus S (PVS) coat protein gene was isolated and the effect of this region on the translation of a downstream open reading frame analysed in vivo. Translation was monitored using the reporter genes B-glucuronidase (GUS) and luciferase (LUC). Translational enhancement was assayed transiently using DNA microprojectile bombardment into both leaf and pollen tissue and also by polyethylene glycol mediated transfection of tobacco protoplasts. In both cases the presence of this region resulted in a 2-3 fold increase in translation when compared to reporter expression with synthetic leader and authentic plant leader constructs. Tobacco plants stability transformed with this PVS 101 nucleotide region and downstream GUS gene gave 4 times the level of translation over synthetic leader GUS control plants.
Botany Department, University of Leicester, U.K.
R
R
Turner
N
N
Bate
D
D
Twell
G D
GD
Foster
D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics*; D017788:Carlavirus / Q000235:genetics* / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005966:Glucuronidase / Q000096:biosynthesis; D008156:Luciferases / Q000096:biosynthesis; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010947:Plants, Toxic; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000096:biosynthesis; D011523:Protoplasts; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D011198:Solanum tuberosum / Q000821:virology; D014026:Nicotiana; D014162:Transfection
D004279:DNA, Viral; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D012367:RNA, Viral; D011994:Recombinant Proteins; D008156:Luciferases; D005966:Glucuronidase