The gene for the major cuticular wax-associated protein and three homologous genes from broccoli (Brassica oleracea) and their expression patterns.
The gene for the major protein (WAX9) found in surface wax of broccoli, designated wax9D, and three homologous genes (wax9A, B and C) were isolated from a genomic library using the previously isolated cDNA encoding the WAX9 protein as the probe; all four genes were sequenced. Genomic Southern blot analysis using the WAX9 cDNA as a probe showed the presence of at least four homologous genes in broccoli genome. The sequence of the originally isolated WAX9 cDNA matched with that of gene D. All four genes have an intron two codons before the stop codon. The putative promoter regions of the four genes, beyond the first 200 bp immediately 5' to the translation start sites, are quite different. Essential elements such as TATA and CAAT boxes and several regions homologous to the promoter regions of other plant Itp genes were identified. The expression patterns of the genes were determined by RT-PCR with gene-specific primers and sequencing of the PCR products. All the genes were expressed in leaves and flower buds. While genes A, B and D also were expressed in stems and open flowers, expression of gene C was not detected in these organs. None of them were expressed in roots. The 972 bp 5'-flanking region of wax9D when fused to beta-glucuronidase (GUS) gene, directed tissue-specific GUS expression in transgenic tobacco plants; GUS expression was found in the epidermis of leaves, stems and flower petals, sepals, trichomes, and ovules but not in roots.
Pyee
J
J
Ohio State Biotechnology Center, Columbus.
Kolattukudy
P E
PE
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D002352:Carrier Proteins; D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C087488:WAX9 protein, Brassica oleracea; D005966:Glucuronidase