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Molecular Biology and Gene Regulation

New Methods for Extraction and Quantitation of Zeins Reveal a High Content of -Zein in Modified opaque-2 Maize ¹

Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana 47907, National Corn and Sorghum Research Center, CNPMS/EMBRAPA, Brazil, Department of Plant Science, University of Arizona, Tucson, Arizona 85721

We have developed methods for quantitative extraction and analysis of zeins from maize (Zea mays L.) flour. Extraction involved solubilization of total endosperm proteins in an alkaline buffer containing SDS and 2-mercaptoethanol with subsequent precipitation of nonzein proteins by the addition of ethanol to 70%. Analysis of these proteins by SDS-PAGE with Coomassie blue staining and by Western blotting and ELISA assay with zein antibodies revealed that this extraction method is more quantitative than the traditional Landry-Moureaux procedure, especially for the - and -zeins. This method was used to extract and analyze the zein content of several `Quality Protein Maize' (QPM) varieties developed by the International Maize and Wheat Improvement Center. QPM varieties contain `modifier genes' that confer a vitreous phenotype on opaque-2 genotypes, while maintaining the elevated levels of lysine and tryptophan characteristic of this mutant. This analysis revealed that the QPM types contain 2 to 4 times the amount of the -zein than unmodified opaque-2 or normal maize varieties. Possible relationships between the high expression of the -zein and the modified opaque phenotype are discussed.

³ Recipient of a graduate fellowship from CIMMYT.

⁴ Recipient of a Rockefeller Foundation fellowship.

¹ This work was supported by NIH grant GM 36970-03 to B. A. L. This is journal paper No. 12044 of the Purdue University Agricultural Experiment Station.

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