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Isolation and Characterization of Two Promoters from Linseed for Genetic Engineering

^a National Research Council of Canada, Plant Biotechnology Institute, Saskatoon, SK, Canada S7N 0W9
^b Crop Development Centre, Univ. of Saskatchewan, Saskatoon, SK, Canada S7N 5A8

View larger version (55K): [in a new window]   Fig. 1 Partial restriction map of the SAD1 gene (a) and identification of SAD1 and SAD2 genes and their promoters by DNA blot analysis (b). E, exon; I, intron; and UT, untranslated regions

View larger version (44K): [in a new window]   Fig. 2 Nucleotide sequence of the 5' regulatory regions of the two SAD genes. Homologous nucleotides (nt) are represented by a dash (-), gaps by a dot (.), and additions by lower case letters. A putative transcriptional site is indicated by +1, and a TATA box is overlined. Key restriction sites are also shown

View larger version (28K): [in a new window]   Fig. 3 Expression of a heterologous gene, uidA, by the two SAD gene promoters in various tissues of linseed. Different tissues are abbreviated as YL+A, young leaves and apices; ML, mature leaves; S, stems; R, roots; B, buds; 12 OF, half open flowers; Fl, fully open flowers; and MS, seeds at about mid-development. The data presented are means of assays conducted on T1 progeny of two plants transformed with a tandem 35S promoter (2x35S) construct, T1 and T3 progeny of two plants transformed with pCDC214 (SAD1) construct, and T1 progeny of two plants transformed with pCDC220 (SAD2) construct

View larger version (13K): [in a new window]   Fig. 4 Expression of a heterologous gene, uidA, by the two SAD gene promoters during linseed seed development and in relation to fatty acid and protein biosyntheses. For GUS assays, the data represent means of assays done on T1 progeny of two plants transformed with a tandem 35S promoter (2x35S) construct, T1 and T3 progeny of two plants transformed with pCDC214 (SAD1) construct, and T1 and T3 progeny of a plant transformed with pCDC220 (SAD2) construct. For fatty acids, three individual embryos of linseed cv. McGregor were analyzed at various stages. For protein content, data are means of estimations done on two transgenic plants transformed with pCDC214 and 220