Cereal Chem 61:141 - 146. | VIEW
Improved Two-Dimensional Electrophoretic Separation of Zein Proteins: Application to Study of Zein Inheritance in Corn Genotypes.
J. S. Wall, D. A. Fey, J. W. Paulis, and J. Landry. Copyright 1984 by the American Association of Cereal Chemists, Inc.
The heterogeneity and genetic variability of polypeptides constituting the protein zein of corn (Zea mays L.) were demonstrated by two-dimentional polyacrylamide gel electrophoresis consisting of isoelectric focusing in the first direction and migration in aluminum lactate pH 3.5 buffer in the second. Zein was extracted from ground corn endosperm with 70% ethanol; the protein was reduced and its sulfhydryls alkylated. Isoelectric focusing was performed in thin gel slabs containing Ampholines (pH 6-8) in 6 M urea at 4 C with contant power. Strips containing resolved zeins were cut from gel slabs, equilibrated with second-dimension buffer, and then inserted into slots in pre-electrophoresed aluminum lactate (pH 3.5) 8M urea gel slabs. After electrophoresis at 4 C and constant power, the 25-40 different zein polypeptides were made visible by silver staining. Zeins from extracts of different inbreds have some different components. Electrophoretic patterns of zeins from hybrids contained polypeptides from both parents, but the female's contribution was quantitatively greater. Inbreds that were derived from crosses usually contained some zeins from both parents. Electrophoretic patterns can characterize zeins in corn inbreds and can aid in relating corn hybrids and inbreds to parental lines.