RUI: Mutational Analysis of Gene Regulation in Maize

阿勒曼9603747 r是玉米中通过其产物螺旋-环-螺旋转录因子的功能调节花青素生物合成途径中基因表达的四个基因座之一。 r决定发育过程中花色素苷沉积的时间和组织特异性。 通过分析影响单转录单位r等位基因调控的突变体，主要研究者为研究r的组织特异性奠定了基础。 R-st复合体的R（Sc）基因指定了籽粒的糊粉层以及幼苗的胚芽鞘和盾片节中的色素沉着。 正在进行的研究已经鉴定了一个493 bp的序列（B' not），其位于Sc调节区中的推定增强子元件中。 该片段可以在某些异等位基因组合中通过拟议的基因转换事件（具有移位的近端区域）定期缺失，以产生增强的Sc表达Sc| P（增强）。 或者，可以通过转座因子Ds诱变来破坏B ′ not元件或周围的增强子区。 Ds插入在这个区域内的色素沉着时，他们在一个方向或完全时，他们在其他方向，可能是通过破坏正常的域边界。上游DNA序列Ds插入突变体，它们的衍生物，和一组选定的三个变异等位基因代表扰动强种子颜色等位基因（Sc），以确定一类r等位基因的结构和功能之间的关系。 通过DNA序列分析，将确定Sc| P（增强的）衍生物生产是通过基因转化。 还将分析色素被条件性阻断的一类等位基因的染色质结构。 将使用表型测定和最终DNA凝胶印迹分析筛选仅r等位基因不同的玉米小种的已建立的集合，以定位其他相关的Sc型等位基因。 基因筛选将是一个区域，以便定位参与发育基因表达的其他区域。 这些研究构成了控制高等植物基因表达的调控序列的详细遗传解剖，对于理解植物发育过程中基因是如何调控的具有根本的重要性。 通过使用诱变，其中起始材料是一系列变异等位基因和单个转录单位，r等位基因决定相同组织中的色素沉着。 鉴定玉米中的组织特异性调控元件对于基因工程这一重要农艺物种至关重要。

Alleman 9603747 r is one of four loci in maize that regulates the expression of the genes in the anthocyanin biosynthetic pathway through the function of its product, helix-loop-helix transcription factor. r determines the timing and tissue-specificity of anthocyanin deposition during development. By analyzing mutants affecting the regulation of single transcriptonal unit r alleles, the principal investigator has established the basis for investigating tissue-specificity of r. The R(Sc) gene of the R-st complex specifies pigmentation in the aleurone layer of the kernel and the coleoptile and scutellar node of the young seedling. On going studies have identified a 493 bp sequence (B' not) which is located in putative enhancer element in the Sc regulatory region. This segment can be regularly deleted in certain heteroallelic combinations via a proposed gene conversion event with a displaced proximal region to produce enhanced Sc expression Sc|P(enhanced) . Alternatively, the B'not element or the surrounding enhancer region can be disrupted by transposable element Ds mutagenesis. Ds insertions within this region block pigmentation slightly when they are in one orientation or totally when they are in the other orientation, possibly by disrupting the normal domain boundary. The upstream DNA sequence Ds insertional mutants, their derivatives, and a selected group of three variant alleles representing perturbation in strong seed color alleles (Sc) to determine the relationship between the structure and function of one class of r alleles. Using this DNA sequence analysis, it will be determined the mechanism for Sc|P(enhanced) derivative production is by gene conversion. Chromatin structure will also be analyzed for a class of alleles for which pigment is conditionally blocked. An established collection of races of maize differing only by the r allele will be screened using a phenotypic assay and eventually DNA gel blot analysis to locate other related Sc-type alleles. A genetic screen will be region in order to localize other regions that are involved in developmental gene expression. The studies constitute a detailed genetic dissection of the regulatory sequences controlling gene expression in higher plants and are of fundamental importance for understanding how genes are regulated during plant development. By using mutagenesis in which the starting material is a series of variant alleles and single transcriptional unit r alleles determining pigmentation in the same that tissue. The identification of tissue-specific regulatory elements in maize is crucial in order to genetically engineer this agronomically important species.