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Analysing the function of existing and novel genetic promoters for tissue-specific expression of transgenes in Zea mays

Deliverables

This result relates to the major objective of the project, the isolation and characterization of promoter motifs conferring tissue specificity on the expression of associated genes in maize. The tissue targets are leaf senescence, kernel development, pollen and seedling development, pathogenesis, and constitutive. Approximately 40 new promoter sequences have been isolated and sequenced, and over half this number have been further characterized by GUS, Mu insertion or deletion analysis. They include: -5 senescence, at least 2 showing Mu insertions, 3 analysed by GUS, 1 showing developmentally specific action in another monocot. -7 MADS box promoters, all analysed by transient expression in the Opaque-2 based system. -3 histone deacetylase complex promoters, transformed into maize with GUS. -10 pollen-specific, 2 of which transformed with GUS into maize. -2-seedling-specific. -7 constitutive, two of which have been transformed with GUS into maize. -4 pathogen-induced, transformed into maize with GUS. -2 guard cell-specific, 1 transformed with GUS. The promoters may be used for generation of GM maize with time and tissue specific expression of agronomically and industrially useful traits, leading to development of new varieties by commercial seed companies. 1. The result is a set of products, namely, a range of genetic promoters that extend the options for controlled expression of new genes introduced to maize in a tissue- or development-specific fashion. 2. The result is relevant to the field of Biotechnology, with the most immediate potential applications likely to be in Agriculture. 3. The number of characterised promoters available for manipulation of monocot crops is restricted; the present result significantly increases the range of such elements available to plant biotechnology. 4. Methods used to identify and analyse the promoters described in this results are amongst the best currently available and include efficient procedures for EST generation, PCR-based cloning, reverse genetics using maize mutator, high-output maize transformation procedures and transient expression analysis. 5. Exploitation of this result will rely on a range of IP-protected technologies related to transformation methodology and would require licensing or other agreements to proceed beyond pre-competitive R&D.