Isolation and nucleotide sequences of cDNA clones encoding ADP-glucose pyrophosphorylase polypeptides from wheat leaf and endosperm |
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Authors: | Mark R Olive R John Ellis Wolfgang W Schuch |
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Institution: | (1) Plant Biotechnology Group, I.C.I. Seeds, P.O. Box 11 The Heath, WA7 4QE Runcorn, Cheshire, United Kingdom;(2) Present address: Division of Plant Industry, C.S.I.R.O., G.P.O. Box 1600, 2601 Canberra, Australia;(3) Department of Biological Sciences, University of Warwick, CV4 7AL Coventry, United Kingdom |
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Abstract: | A cDNA clone (WL : AGA.1) encoding wheat leaf ADP-glucose pyrophosphorylase has been isolated from a gt11 expression library, by immunological screening with anti-spinach leaf ADP-glucose pyrophosphorylase serum. The WL : AGA.1 cDNA is 948 bp long and contains approximately 55% of the complete wheat leaf ADP-glucose pyrophosphorylase mRNA sequence, estimated from Northern blot experiments. A wheat endosperm cDNA library was subsequently constructed in gt11 and six clones hybridising to the cDNA insert of clone WL : AGA.1 were isolated. The longest of these wheat endosperm ADP-glucose pyrophosphorylase cDNAs, clone WE : AGA.7, is nearly full-length (1798 bp), indicated by Northern blot analysis of wheat endosperm mRNA and nucleotide sequence analysis.Southern hybridisation analysis and restriction enzyme mapping indicated that the wheat leaf and wheat endosperm ADP-glucose pyrophosphorylase cDNAs and genes are members of two distinct gene families. In addition, restriction enzyme mapping revealed polymorphism in the wheat endosperm ADP-glucose pyrophosphorylase cDNAs, indicating the existence of at least two wheat endosperm ADP-glucose pyrophosphorylase gene sub-families.Subsequent nucleotide sequence analysis indicates that there is approximately 55% identity between wheat leaf and wheat endosperm ADP-glucose pyrophosphorylase cDNAs. In contrast, members of each sub-family of endosperm cDNA, represented by clones WE : AGA.3 and WE : AGA.7, are 96% identical. |
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Keywords: | ADP-glucose pyrophosphorylase cDNAs gene organisation polypeptides wheat |
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