CELL REGULATION--BIOCHEMICALLY ISOLATED DNA SEGMENTS

描述(改编自《调查员摘要》)：调查员 建议对酵母菌和拟南芥进行全基因组分析。他们 将广泛利用酵母的基因组序列。他们会发展成 并应用允许并行分析所有区域的新技术 基因组。他们将参加一项国际合作，删除 酵母基因组中的每一个基因。他们将应用一种新开发的 结合了DNA序列标签和相应的DNA芯片的技术 来自Affymetrix的，将允许对每个 并行删除。研究人员还将构建一种微阵列 含有每个已鉴定的开放阅读框的聚合酶链式反应产物的玻璃 以及酵母基因组的每个基因间隔区。这些微阵列将被 用于并行测量每个基因在许多情况下的表达水平 国家，绘制DNA复制的起源图，并遵循复制分叉 细胞周期中的运动。他们将对一个集合进行分离和排序 整个基因组中的10,000个DNA结合位点，并鉴定 相应的DNA结合蛋白。基因组中的每一个基因都将被克隆 作为聚合酶链式反应产物，并用独特的20聚体集合进行分子标记 序列以允许并行屏幕和选择。这些产品的一次使用 试剂将用于开发一种药物筛选程序，允许所有基因 在基因组中被平行靶向。调查人员还将 拟南芥高分辨率双等位基因标记遗传图谱的构建。 复杂性状的遗传作图将通过多重聚合酶链式反应和 芯片杂交，以实现快速和自动化分析。

DESCRIPTION (Adapted from the Investigator's Abstract): The investigators propose to conduct whole genome analysis on yeast and A. thaliana. They will make extensive use of the genome sequence of yeast. They will develop and apply new technology that allows parallel analysis of all regions of the genome. They will participate in an international collaboration to delete every gene in the genome of yeast. They will apply a newly developed technology that incorporates a DNA sequence tag and a corresponding DNA chip from Affymetrix that will allow quantitative phenotypic analysis of every deletion in parallel. The investigators will also construct a microarray on glass that contains the PCR products of every identified open reading frame and every intergenic region of the yeast genome. These microarrays will be used to measure in parallel expression levels of every gene under many states, map the origins of DNA replication and follow replication fork movement during the cell cycle. They will isolate and sequence a collection of 10,000 DNA binding sites throughout the genome and identify the corresponding DNA binding protein. Every gene in the genome will be cloned as a PCR product and molecularly tagged with the collection of unique 20mer sequence to allow parallel screens and selections. One use of these reagents will be for developing a drug screen program that allows all genes in the genome to be targeted in parallel. The investigators will also develop a high resolution biallelic marker genetic map for Arabidopsis. Genetic mapping of complex traits will be conducted by multiplexed PCR and chip hybridization to allow rapid and automated analysis.