The construction and testing of maize transcriptional fusions in yeast (Saccharomyces cerevisiae)

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Virginia Tech


The specific goal of this study was to construct and test transcriptional fusions of zein promoters and a yeast reporter gene that will serve as part of a two plasmid system that will allow for the identification of maize transcriptional regulators of zein genes. Zein genes are expressed coordinately and tempO~ly during endosperm development and are controlled at the transcriptional level (Pedersen et aL, 1980; Kodryzcki et al. t 1989). The accumulation of zein proteins in the endosperm presents an ideal model system to study plant gene regulation. These proteins are synthesized only in the endosperm tissue, and their concentration in the endosperm determine the nutritional quality of the seed.

Because of the coordinate and temporal regulation of zein gene transcription, there is a strong likelihood that there exists positive regulatory elements of zein gene expression during early endosperm development. We know that the control of storage protein gene expression is mediated by regulatory elements in the endosperm of maize seeds. It has been shown that the recessive mutation opaque-2 (02) specifically reduces the 22,000 zein polypeptide .. Schmidt et at (1990) and Aukennan et at (1991) show that the wild-type 02 encodes a protein containing a basic leucine zipper domain