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United States Department of Agriculture

Agricultural Research Service

Title: Isolation of the Promoter for a Gene Preferentially Expressed in Barley Leema/palea Tissue

Authors
item Sathish, Puthigae - UNIVERSITY OF WISCONSIN
item Federico, Maria - UNIVERSITY OF WISCONSIN
item Kaeppler, Heidi - UNIVERSITY OF WISCONSIN
item Skadsen, Ronald

Submitted to: Congress on In Vitro Biology
Publication Type: Abstract Only
Publication Acceptance Date: June 1, 1999
Publication Date: N/A

Technical Abstract: We have isolated several genes that are expressed preferentially in the lemma and palea floret tissues of barley (Hordeum vulgare L.). The 3' regions of the genes were initially isolated using the differential display technique. We proceeded to analyze the expression of one such gene and isolate its promoter. Using RNA blot analysis, the corresponding mRNA was found to be approximately 700 bases long. This gene is expressed in the lemma/palea but no in the developing endosperm, roots or etiolated shoots. A single genomic clone for this gene was obtained from a screening of a barley BAC library (courtesy of Dr. A. Kleinhofs, Washington State University. After restriction map analysis, a 3 kb fragment was cloned in pBluescript S/K (Stratagene) and sequenced to characterize the upstream region. We determined the 5' regulatory region necessary for high level transient expression of a green fluorescent protein (gfp) reporter gene in the lemma and palea. This was accomplished by constructing plasmids containing progressive 5' deletions of the gene and fusing them, in frame, to the coding sequences of a gfp with the nopaline synthase (nos) terminator. We present the results from the homology analysis of the gene and transient expression pattern of the gfp under the 5' regulatory region of this gene. This promoter may be useful to direct the expression of anti-fungal genes against Fusarium graminearum, which causes fusarium head blight of barley and wheat.

Last Modified: 12/22/2014
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