Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/236
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Type: Journal article
Title: Biochemical evidence linking a putative callose synthase gene with (1→3)-β-d-glucan biosynthesis in barley
Other Titles: Biochemical evidence linking a putative callose synthase gene with (1->3)-beta-d-glucan biosynthesis in barley
Author: Li, J.
Burton, R.
Harvey, A.
Hrmova, M.
Wardak, A.
Stone, B.
Fincher, G.
Citation: Plant Molecular Biology: An International Journal on Molecular Biology, Molecular Genetics and Biochemistry, 2003; 53(1):213-225
Publisher: Kluwer Academic Publ
Issue Date: 2003
ISSN: 0167-4412
1573-5028
Statement of
Responsibility: 
Jing Li, Rachel A. Burton, Andrew J. Harvey, Maria Hrmova, Ahmad Z. Wardak, Bruce A. Stone and Geoffrey B. Fincher
Abstract: A putative barley (1→3)-β-D-glucan synthase cDNA of 6.1 kb, which is homologous to the yeast FKS gene, was assembled from DNA fragments obtained through screening of barley cDNA and BAC libraries, and by PCR amplification. The corresponding gene, designated HvGSL1, is a member of a family of at least six genes in barley. Gene transcripts are detected at relatively high levels in early developing grain, florets, coleoptiles and roots, but not in leaves infected with a fungal pathogen. A (1→3)-β-D-glucan synthase has been purified more than 60-fold from barley suspension-cultured cells by detergent extraction, CaCl2 treatment, sucrose density gradient centrifugation and non-denaturing gel electrophoresis. The enzyme synthesizes (1→3)-β-D-glucan in vitro and is recognized by antibodies raised against a 17 kDa protein generated by heterologous expression of a fragment of the HvGSL1 cDNA. Furthermore, mass spectrometric analyses show that tryptic peptides produced by in-gel digestion of the active enzyme match peptides predicted from the gene sequence. Thus, the amino acid sequence predicted from the HvGSL1 gene has been linked with the actual amino acid sequence of an active (1→3)-β-D-glucan synthase fraction from barley.
Keywords: Active site
gene expression
genetic mapping
GSL
Hordeum vulgare
purification
Description: The original publication can be found at www.springerlink.com
Rights: © 2003 Kluwer Academic Publishers
DOI: 10.1023/B:PLAN.0000009289.50285.52
Published version: http://dx.doi.org/10.1023/b:plan.0000009289.50285.52
Appears in Collections:Agriculture, Food and Wine publications
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