DEVELOPMENTAL GENETICS OF AN INDUCIBLE GENE SYSTEM

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On the basis of our previous mutant analyses, we know that particular regions near Adh1 exons can affect 3' processing, quantitative and organ-specific regulation of ADH1 message expression. In the process of using transposons as mutagens, we have discovered two new, useful transposons -- Mu1 and copia-like Bs1 -- and expect that other of our new Adh mutants carry additional new transposons. We now know that the Adhs are co-regulated with a sucrose synthase, phosphogluco isomerase, aldolase and pyruvate decarboxylase; apparently, the anaerobic response involves the synthesis of messages that encode the glycolytic enzymes. I propose to focus on the induction and characterization of organ-specific Adh1 and Sh1 mutants, and of mutants involving anaerobic metabolism directly. In particular, we hope to identify mutants that change the expression of more than one anaerobic protein, expression of organ-specificity of anaerobic protein message synthesis, or of the mechanism that senses oxygen tension. Mutants will be mapped to complementation group, and analyzed for differences in 2-D gel pattern of 35S-Met incorporation into anaerobic proteins; they will be cloned by a double-Mu-clone-pool method of transposon marking if we do not already have a probe among our cDNAs. We also propose to reinvestigate compensatory regulation of Adh1, beginning with a compensated deletion mutant of Adh1. Further, we propose to apply our methods of cell immobilization, current microinjection (iontophoresis) and Mu1-vector construction to transforming maize cells in culture.