Simple ELISA Detection of a New Polymorphic Ha Locus Encoded Protein |
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| Institution: | 1. Department of Chemistry, University of Zabol, Zabol, Iran;2. Department of Nanotechnology and Advanced Materials, Materials and Energy Research Center, Karaj, Iran;1. Stone Unit, Guy''s and St. Thomas’ NHS Foundation Trust, London, UK;2. Stone / EndoUrology Unit, University College Hospital, London, UK;1. Lomonosov Moscow State University, 119991 Moscow, Russia;2. Pirogov Russian National Research Medical University, 117997 Moscow, Russia;3. Koltzov Institute of Developmental Biology of Russian Academy of Sciences, 119334 Moscow, Russia;4. Institute of Biomedical Problems of Russian Academy of Sciences, 123007 Moscow, Russia;1. Department of Organic Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097 Warsaw, Poland;2. Department of Inorganic and Analytical Chemistry, University of Szeged, Dóm tér 7, H-6720 Szeged, Hungary;1. School of Chemistry, The University of Sydney, Sydney, NSW 2006, Australia;2. Department of Chemistry and the Cell and Molecular Biology Program, Colorado State University, Fort Collins, CO 80523, USA |
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| Abstract: | A rapid two-site sandwich ELISA was developed for detection of a previously uncharacterised protein encoded at the Ha locus on chromosome 5DS of wheat (Triticum aestivum). The assay used the combined specificity of two antibodies to detect a protein that was soluble in aqueous alcohol, salt solutions and water. It was expressed in the endosperm of all soft wheats and Triticum tauschii accessions tested. The ELISA was highly specific, with no signal obtained with varieties that did not express the protein. The presence of the 5DS-encoded protein correlated with a significant change in both water absorption and average hardness and particle size indices in a doubled haploid population derived from a cross between cvs. Cranbrook×Halberd. Only some hard varieties expressed this protein indicating that the protein is not predictive for hardness. However, it may be a new factor, or a marker for a new factor, affecting kernel texture. A polypeptide ofMr 66 000 was purified from an extract of Halberd flour by immunoaffinity chromatography. Its N-terminal amino acid sequence identified it as an albumin with high homology to both mammalian serum albumins and sucrose synthase from a range of cereals. The assay may be valuable in wheat breeding programmes for assessing kernel texture where the parents are of different ELISA phenotype, or for varietal identification, as the expression of the polypeptide is variable in hard wheat varieties. |
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