Subdomains of the octopine synthase upstream activating element direct cell-specific expression in transgenic tobacco plants.
Previous work has shown that the octopine synthase (ocs) gene encoded by the Agrobacterium tumefaciens Ti-plasmid contains an upstream activating sequence necessary for its expression in plant cells. This sequence is composed of an essential 16-bp palindrome and flanking sequences that modulate the level of expression of the ocs promoter in transgenic tobacco calli. In this study, we have used RNA gel blot analysis of RNA extracted from transgenic tobacco plants to show that the octopine synthase gene is not constitutively expressed in all plant tissues and organs. This tissue-specific pattern of expression is determined, to a large extent, by the 16-bp palindrome. Histochemical analysis, using an ocs-lacZ fusion gene, has indicated that the 16-bp palindrome directs the expression of the ocs promoter in specific cell types in the leaves, stems, and roots of transgenic tobacco plants. This expression is especially strong in the vascular tissue of the leaves, leaf mesophyll cells, leaf and stem guard cells, and the meristematic regions of the shoots and roots. Sequences surrounding the palindrome in the upstream activating sequence restrict the expression of the ocs promoter to fewer cell types, resulting in a reduced level of expression of beta-galactosidase activity in the central vascular tissue of leaves, certain types of leaf trichomes, and the leaf primordia.
Kononowicz
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Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Wang
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Habeck
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Gelvin
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SB
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D015152:Blotting, Northern; D004252:DNA Mutational Analysis; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006651:Histocytochemistry / Q000379:methods; D007763:Lac Operon / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D010947:Plants, Toxic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D014018:Tissue Distribution; D014026:Nicotiana / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D000594:Amino Acid Oxidoreductases; C021903:D-octopine dehydrogenase