Isolation of a ubiquitin-ribosomal protein gene (ubi3) from potato and expression of its promoter in transgenic plants.
A genomic clone encoding the potato homolog of the yeast ubiquitin-ribosomal protein fusion gene ubi3 was isolated and characterized. Chimeric genes containing the ubi3 promoter (920 bp of 5' to the ubiquitin start codon) were constructed in which the reporter gene beta-glucuronidase (GUS) was either fused directly to the promoter, or introduced as a translational fusion to the ubiquitin-coding region. After introduction into the potato by Agrobacterium-mediated transformation, GUS activities were measured in leaves and in tubers of transgenic clones. GUS activity was 5- to 10-fold higher in clones expressing the ubiquitin-GUS translational fusion than in clones containing GUS fused directly to the ubi3 promoter. For both types of constructs, GUS activity was highest in meristematic leaves and declined during leaf expansion, then rose again to near the meristematic levels during senescence. GUS activity in tubers was similar to that in young leaves. In contrast to the native ubi3 genes, the chimeric ubi3-GUS transgenes were not activated in the tuber by wounding.
Garbarino
J E
JE
United States Department of Agriculture, Albany, CA 94710.
Belknap
W R
WR
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004247:DNA; D005091:Exons; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012269:Ribosomal Proteins / Q000096:biosynthesis / Q000235:genetics*; D011198:Solanum tuberosum / Q000235:genetics*; D014452:Ubiquitins / Q000096:biosynthesis / Q000235:genetics*
D010940:Plant Proteins; D012269:Ribosomal Proteins; D014452:Ubiquitins; C085995:ubi3 protein, Solanum tuberosum; D004247:DNA; D005966:Glucuronidase