Construction and evaluation of a maize (Zea mays) chimaeric promoter with activity in kernel endosperm and embryo

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2009-03-01
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Shepherd, Colin
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Scott, M. Paul
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Agronomy

The Department of Agronomy seeks to teach the study of the farm-field, its crops, and its science and management. It originally consisted of three sub-departments to do this: Soils, Farm-Crops, and Agricultural Engineering (which became its own department in 1907). Today, the department teaches crop sciences and breeding, soil sciences, meteorology, agroecology, and biotechnology.

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The Department of Agronomy was formed in 1902. From 1917 to 1935 it was known as the Department of Farm Crops and Soils.

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1902–present

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  • Department of Farm Crops and Soils (1917–1935)

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Abstract

Chimaeric promoters contain DNA sequences from different promoters. Chimaeric promoters are developed to increase the level of recombinant protein expression, to precisely control transgene activity or to combat homology-based gene silencing. Sets of chimaeric promoters, each containing different lengths of DNA from maize (Zea mays) 27zn (27 kDa γ-zein) endosperm-specific promoter and the Glb1 (Globulin-1) embryo-specific promoter were created and tested in a transient expression assay of GFP (green fluorescent protein). Promoter fragments with the highest activity were combined to create the chimaeric promoter A27znGlb1. In the context of the chimaeric promoter, the selected Glb1 promoter fragment was necessary and sufficient to activate expression in embryo tissue and was functionally equivalent to the native Glb1 promoter. Similarly, the selected 27zn promoter fragment in the chimaeric promoter was necessary and sufficient to activate expression in endosperm tissue and was functionally equivalent to the native 27zn promoter. Maize transgenic plants containing the A27znGlb1 chimaeric promoter fused to GFP were produced to characterize this promoter in vivo. Quantitative reverse-transcriptase PCR was used to determine that the promoter was active in the embryo, endosperm, pericarp and immature leaf tissues. GFP activity in plants containing the chimaeric promoter was not significantly different in endosperm than the activity of GFP fused to the full-length 27zn promoter, nor was it different in embryo from the activity of GFP fused to the full-length Glb1 promoter. Transgene copy numbers were shown to be between 4 and 12 copies in different events.

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This article is published as Shepherd, Colin T. and Scott, M. Paul. (2009), Construction and evaluation of a maize (Zea mays) chimaeric promoter with activity in kernel endosperm and embryo. Biotechnology and Applied Biochemistry, 52: 233–243, doi:10.1042/BA20070269.

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