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The effect of different promoter-sequences on transient expression of gus reporter gene in cultured barley (Hordeum vulgare L.) cells
Authors:Ravindra N Chibbar  Kutty K Kartha  Raju S S Datla  Nick Leung  Karen Caswell  Clifford S Mallard  Lee Steinhauer
Institution:(1) Plant Biotechnology Institute, National Research Council Canada, 110 Gymnasium Place, S7N 0W9 Saskatoon, Saskatchewan, Canada
Abstract:The cauliflower mosaic virus 35S (35S) and the enhanced 35S (E35S) promoters fused with maize alcohol dehydrogenase (Adh1) intron1 or maize shrunken locus (sh1) intronl along with maize Adh1 and rice actin (Act1) promoters fused to their respective first introns were tested for transient expression of the E.coli beta-glucuronidase (gus) reporter gene in cultured barley (Hordeum vulgare L) cells. The plasmids, carrying the respective promoterintron combinations to drive the gus fused to nopaline synthase (nos) terminator, were introduced into cultured barley cells using a particle gun. The rice Act1 promoter with its first intron gave the highest expression of all promoter intron combinations studied. This was followed by the E35S promoter and no significant differences were observed between the other two promoters tested. The rice actin promoter is now being used to drive selectable marker genes to obtain stably transformed cereal cells.NRCC No. 36482
Keywords:particle bombardment  Hordeum vulgare L    promoter screening  transient expression  gus  cereal transformation
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