The 5’-region of the chitinase gene cabch29, derived from Brassica oleracea var. capitata, has been sequenced and analyzed for cis-acting elements important in controlling gene expression in transgenic tobacco plants...The 5’-region of the chitinase gene cabch29, derived from Brassica oleracea var. capitata, has been sequenced and analyzed for cis-acting elements important in controlling gene expression in transgenic tobacco plants. Different 5’-deletion fragments were linked to reporter geneβ-glucuronidase (GUS) as translational fusions, and the expression of these chimeric genes was analyzed in vegetative organs and tissues. Sequences up to -651 showed some basal GUS activity with nearly equal levels in wounded and intact tissues. The addition of further upstream sequences (-651 to -1284) enhanced expression level, and the expression driven by this fragment was inducible by a factor of two to three-fold by wounding. Histochemical analysis of different tissue from transgenic plants that contain cabch29 promoter-GUS fusion gene demonstrated woundinducible and tissue-specific cabch29 promoter activity in plants containing the 1308 base pair fragment. The location of GUS activity appears to be cell-specific, being highest in vascular cells and epidermal cells of stem, leaf and roots. Meanwhile, the temporal and spatial expression of cabch29-GUS fusion gene has been investigated. Among the different vegetative organs, a high level of GUS activity was observed in stem and a moderate one in roots;whereas, wounding stress led to a high level of GUS in stem and moderate one in leaf.展开更多
文摘The 5’-region of the chitinase gene cabch29, derived from Brassica oleracea var. capitata, has been sequenced and analyzed for cis-acting elements important in controlling gene expression in transgenic tobacco plants. Different 5’-deletion fragments were linked to reporter geneβ-glucuronidase (GUS) as translational fusions, and the expression of these chimeric genes was analyzed in vegetative organs and tissues. Sequences up to -651 showed some basal GUS activity with nearly equal levels in wounded and intact tissues. The addition of further upstream sequences (-651 to -1284) enhanced expression level, and the expression driven by this fragment was inducible by a factor of two to three-fold by wounding. Histochemical analysis of different tissue from transgenic plants that contain cabch29 promoter-GUS fusion gene demonstrated woundinducible and tissue-specific cabch29 promoter activity in plants containing the 1308 base pair fragment. The location of GUS activity appears to be cell-specific, being highest in vascular cells and epidermal cells of stem, leaf and roots. Meanwhile, the temporal and spatial expression of cabch29-GUS fusion gene has been investigated. Among the different vegetative organs, a high level of GUS activity was observed in stem and a moderate one in roots;whereas, wounding stress led to a high level of GUS in stem and moderate one in leaf.
