Abstract
Monoclonal antibodies to hordein polypeptides have been produced and characterized. All antibodies reacted with more than one polypeptide, indicating that different hordein polypeptides have common epitopes. Of the seventeen isolated hybridoma lines, six secreted antibodies which reacted both with B- and C-hordein, five recognized only B-hordein and one secreted antibodies specific for C-hordein polypeptides. Two clones produced antibodies to an unidentified polypeptide doublet with an apparent molecular weight of 43 kD. These two polypeptides have at least one epitope in common with B-hordein polypeptides and are not extractable in water but in alcohol. They are not visible by Coomassie blue staining.
The mutant M56 fails to synthesize B-hordein polypeptides due to a major deletion. It contains polypeptides in the B-hordein region of the gel, which have epitopes recognized by the B-hordein antibodies, but are different from B-hordeins. The 43 kD hordein doublet was more abundant in the mutant than in the wild-type endosperm, suggesting that these polypeptides like the C-hordein polypeptides are synthesized in larger amounts as a consequence of the absent B-hordein polypeptide synthesis. Mutant 1508 which has an impaired synthesis of both B- and C-hordein polypeptides was also found to be defective in the synthesis of the 43 kD polypeptide doublet. It was shown that immunofluorescent techniques may be of practical importance to identify mutants with elevated or reduced amounts of specific hordein polypeptides. Due to the higher sensitivity compared to SDS-PAGE, immunoblotting can be a useful tool for genotype identification in breeding programmes.
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Abbreviations
- CM:
-
Carlsberg Monoclonal
- FITC:
-
fluorescein isothiocyanate
- Ig:
-
immunoglobulin
- kD:
-
kilodalton
- MAb:
-
monoclonal antibody
- p:
-
protein
- PAGE:
-
polyacrylamide gel electrophoresis
- SDS:
-
Sodium dodecyl sulphate
- Tris:
-
tris-(hydroxymethyl)-aminomethane
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Ullrich, S.E., Rasmussen, U., Høyer-Hansen, G. et al. Monoclonal antibodies to hordein polypeptides. Carlsberg Res. Commun. 51, 381–399 (1986). https://doi.org/10.1007/BF02907314
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DOI: https://doi.org/10.1007/BF02907314