Cryptic regulatory elements obtained from plants

C - Chemistry – Metallurgy – 12 – N

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C12N 15/82 (2006.01) A01H 5/00 (2006.01) C12N 5/10 (2006.01) C12N 15/11 (2006.01) C12N 15/63 (2006.01)

Patent

CA 2331842

T-DNA tagging with a promoterless .beta.-glucuronidase (GUS) gene generated transgenic Nicotiana tabacum plants that expressed GUS activity either only in developing seed coats, or constitutively. Cloning and deletion analysis of the GUS fusion revealed that the promoter responsible for seed coat specificity was located in the plant DNA proximal to the GUS gene. Analysis of the region demonstrated that the seed coat-specificity of GUS expression in this transgenic plant resulted from T-DNA insertion next to a cryptic promoter. This promoter is useful in controlling the expression of genes to the developing seed coat in plant seeds. Similarly, cloning and characterization of the cryptic constitutive promoter revealed the occurrence of several cryptic regulatory regions. These regions include promoter, negative regulatory elements, transcriptional enhancers, core promoter regions, and translational enhancers and other regulatory elements.

Le marquage de l'ADN-T par un gène bêta-glucuronidase sans promoteur (GUS) a généré des plantes transgéniques appelées Nicotiana tabacum qui ont exprimé une activité GUS soit uniquement par développement de téguments, soit de manière constitutive. L'analyse du clonage et de la délétion de la fusion GUS a révélé que le promoteur responsable de la spécificité du tégument était implanté dans l'ADN de la plante à proximité du gène GUS. L'analyse de la région a démontré que la spécificité de l'expression de GUS relative au tégument dans ladite plante transgénique procédait de l'insertion de l'ADN-T à côté d'un promoteur cryptique. Ce promoteur régule efficacement dans des semences l'expression de gènes relativement au tégument en croissance. De même, le clonage et la caractérisation du promoteur cryptique constitutif ont révélé la présence de plusieurs régions régulatrices cryptiques. Ces régions incluent un promoteur, des éléments régulateurs négatifs, des amplificateurs transcriptionnels, des régions promotrices principales et des amplificateurs traductionnels ou autres éléments régulateurs.

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