title: Isolation and characterization of the threonine deaminase promoter in Nicotiana attenuata.
abstract: The enzyme encoded by the threonine deaminase (TD) gene catalyzes the conversion of threonine to α-keto butyrate in the biosynthesis of isoleucine (Ile). In Nicotiana attenuata, TD transcripts accumulate constitutively in cotyledons and flowers and are elicited in leaves by wounding, herbivore attack, and methyl jasmonic acid (MeJA) treatment. To understand TD's unique pattern of expression, we isolated a genomic clone of the TD gene from N. attenuata and characterized its promoter. T2 transgenic plants, each harboring single copies of fusions of different parts of the 5' non-coding region to the β-glucuronidase reporter gene, demonstrated that the promoter was constitutively expressed in seedlings and flowers, and elicited in leaves by wounding or by MeJA treatment. Promoter deletion analysis defined the promoter regions capable of directing minimal expression in cotyledons and anthers as -142 to -31bp, and in stigmas as -289 to -231bp. Regions from -142 to -31bp were found to be important for basal elicitation in leaves by both wounding and MeJA treatment. These promoter elements may prove valuable in biotechnological applications.
authors.affiliation: Department of Molecular Ecology, Max-Planck Institute of Chemical Ecology, Hans Knöll Str. 8, D-07745 Jena, Germany.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Kang
authors.affiliation: Department of Molecular Ecology, Max-Planck Institute of Chemical Ecology, Hans Knöll Str. 8, D-07745 Jena, Germany. Electronic address: baldwin@ice.mpg.de.
authors.forename: Ian T
authors.initials: IT
authors.lastname: Baldwin
keywords: Promoter; Threonine deaminase; Wounding; β-Glucuronidase