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CELL REGULATION--BIOCHEMICALLY ISOLATED DNA SEGMENTS

细胞调节——生化分离的 DNA 片段

基本信息

批准号：
6469447
负责人：
Ronald Wayne Davis
金额：
$ 13.27万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-03-01 至 2002-03-31
项目状态：
已结题

项目摘要

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.

描述（改编自研究者摘要）：研究者建议对酵母和A. thaliana. 他们将广泛利用酵母的基因组序列。他们将开发并应用新技术，允许并行分析的所有地区的基因组他们将参与一项国际合作，酵母基因组中的每一个基因。他们将采用一种新开发的一种结合DNA序列标签和相应DNA芯片的技术这将允许对每一个人进行定量表型分析，平行删除。研究人员还将构建一个微阵列，含有每个已识别开放阅读框的PCR产物的玻璃杯和酵母基因组的每个基因间区域。这些微阵列将用于在许多条件下平行测量每个基因的表达水平，国家，映射DNA复制的起点，并遵循复制叉细胞周期中的运动。他们会分离并排序一个集合基因组中10，000个DNA结合位点，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PCR产物，并用独特的20聚体的集合进行分子标记，序列，以允许并行屏幕和选择。一个使用这些试剂将用于开发一种药物筛选程序，在基因组中被平行靶向。调查人员还将建立拟南芥高分辨率双等位基因标记遗传图谱。复杂性状的遗传作图将通过多重PCR进行，芯片杂交以允许快速和自动化分析。