DEVELOPMENTAL GENETICS OF AN INDUCIBLE GENE SYSTEM

根据我们之前的突变分析，我们知道， Adh 1外显子附近的区域可以影响3'加工，定量和 器官特异性调节ADH 1信息表达。 过程中 利用转座子作为诱变剂，我们发现了两个新的，有用的， 转座子--Mu 1和copia样Bs 1--并期望我们的其他新的 Adh突变体携带额外的新转座子。 我们现在知道Adhs 与蔗糖合成酶、磷酸葡萄糖异构酶、醛缩酶 和丙酮酸脱羧酶;显然，厌氧反应涉及 合成编码糖酵解酶的信息。 我建议把重点放在诱导和表征器官特异性 Adh 1和Sh 1突变体，以及涉及厌氧代谢的突变体 直接. 特别是，我们希望确定突变体，改变 一种以上厌氧蛋白的表达， 厌氧蛋白质信息合成的器官特异性，或 感觉氧气压力的机制。 突变体将被映射到 互补组，并分析在2-D凝胶模式的差异， 35 S-Met掺入厌氧蛋白;它们将通过 转座子标记的双Mu克隆池方法，如果我们还没有 在我们的基因组中做一个探针 我们还建议重新调查补偿性 调节Adh 1，从Adh 1的补偿缺失突变体开始。 此外，我们建议应用我们的细胞固定化方法， 显微注射（离子导入）和Mu 1载体构建以转化 玉米细胞培养。

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.