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GrainGenes Reference Report: JEB-52-243

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Reference	JEB-52-243
Title	Endosperm-specific activity of a storage protein gene promoter in transgenic wheat seed
Journal	Journal of Experimental Botany
Year	2001
Volume	52
Pages	243-250
Author	Lamacchia C [ Show all 9 ]
Abstract	Summary: The characterization of the promoter of a wheat (Triticum aestivum) cv. Cheyenne high molecular weight glutenin subunit (HMW subunit) gene, Glu-1D-1 is reported. The nucleotide sequence of the promoter from position -1191 to -650 with respect to the transcription start site was determined, to add to that already determined. Analysis of this region of the promoter revealed the presence of an additional copy of part of the primary enhancer sequence and sequences related to regulatory elements present in other wheat seed protein genes. A chimaeric gene was constructed comprising the 5' flanking region of the Glu-1D-1 gene from position -1191 to +58, the coding region of the UidA (Gus) gene, and the nopaline synthase (Nos) gene terminator. This chimaeric gene was introduced into wheat (Triticum durum cv. Ofanto) by particle bombardment of inflorescence explants. Two independent transgenic lines were produced, and both showed expression of the Gus gene specifically in the endosperm during mid-development (first detected 10-12 d after anthesis). Histochemical analysis of homozygous T2 seed confirmed this pattern of expression, and showed that expression was initiated first in the central lobes of the starchy endosperm, and then spread throughout the endosperm tissue, while no expression was detected in the aleurone layer. Native HMW subunit protein was detectable by Western analysis 12-14 d after anthesis, consistent with concurrent onset of activity of the native and introduced HMW subunit gene promoters
External Databases	Pubmed: 11283168
Keyword	[ Hide all but 1 of 55 ] aleurone aleurone layer barley bombardment bread wheat characterization cheyenne chromosome-1a durum element elements endosperm enhancer explants gene gene expression gene promoter glutenin glutenin subunit gus high molecular weight high molecular weight glutenin subunit histochemical analysis hmw hordein inflorescence molecular weight molecular-weight glutenin nucleotide nucleotide sequence onset particle bombardment position promoter protein gene protein genes reporter gene seed seed protein sequence spread starchy endosperm storage storage protein subunit synthase tissue transcription transcription start site transgenic transgenic wheat triticum aestivum triticum durum weight wheat seed

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