DEVELOPMENTAL GENETICS OF AN INDUCIBLE GENE SYSTEM

根据我们之前的突变分析，我们知道这个特定的 Adh1外显子附近的区域可影响3‘加工、数量和 ADH1信息表达的器官特异性调节。在…的过程中 利用转座子作为诱变剂，我们发现了两种新的、有用的 转座子--MU1和类似Copia的BS1--并期待我们的另一个新的 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、醛缩酶共同调节 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。 我建议把重点放在器官特异性的诱导和特征上 Adh1和SH1突变体，以及与无氧代谢有关的突变体 直接去吧。特别是，我们希望找出能改变基因的突变体 表达不止一种厌氧蛋白，表达 厌氧蛋白质信息合成的器官特异性，或 感觉氧气紧张的机制。变种人将被映射到 互补组，并分析其二维凝胶图谱的差异 35S-Met掺入厌氧蛋白质中；它们将通过 如果我们还没有转座子标记的双Mu克隆池方法 我们的DNA中的一个探测器。我们还建议重新调查补偿性 Adh1的调节，从Adh1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，目前 微注射(离子导入)与MU1载体构建转化 培养中的玉米细胞。

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.