Biochemical evidence linking a putative callose synthase gene with (1→3)-β-d-glucan biosynthesis in barley
Date
2003
Authors
Li, J.
Burton, R.
Harvey, A.
Hrmova, M.
Wardak, A.
Stone, B.
Fincher, G.
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Advisors
Journal Title
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Volume Title
Type:
Journal article
Citation
Plant Molecular Biology, 2003; 53(1):213-225
Statement of Responsibility
Jing Li, Rachel A. Burton, Andrew J. Harvey, Maria Hrmova, Ahmad Z. Wardak, Bruce A. Stone and Geoffrey B. Fincher
Conference Name
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.
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The original publication can be found at www.springerlink.com
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© 2003 Kluwer Academic Publishers