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SGER: An ORF - Rescue Vector for Crop Genome Sequencing

SGER：用于作物基因组测序的 ORF - 救援载体

基本信息

批准号：
0121417
负责人：
Patrick Schnable
金额：
$ 8.96万
依托单位：
Iowa State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2001
资助国家：
美国
起止时间：
2001-06-01 至 2003-05-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=0121417&HistoricalAwards=false
关键词：
SGER ORF Rescue Vector Crop

项目摘要

ABSTRACTThe maize genome is thought to comprise islands of lightly methylated genes in a sea of highly methylated, repetitive DNA. Complete sequencing and assembly of the whole maize genome is currently not feasible because of the repetitive DNA. Methods are beginning to be developed that will allow the regions of the maize genome that contain the genes to be filtered out for sequencing. It is expected that once validated in maize, such methods could be used for similar sequencing projects in other crop species having complex genomes.A novel expression vector system has been developed to rescue directly open reading frames (ORFs) from genomic DNA. Preliminary data suggest that this "ORF Rescue" vector system is a potentially powerful tool for gene discovery in complex crop genomes. Experiments will be performed to validate the utility of this vector system for preparing filtered template DNA for genome sequencing. The three specific aims of the project are to:1] evaluate the ability of the ORF Rescue vector to filter genes from maize genomic DNA; 2] evaluate the ability of the ORF Rescue vector to filter genes from maize DNA cloned into Bacterial Artificial Chromosomes (BACs); 3] determine whether novel ORFs define monocot-specific genes. This work merits funding as a Small Grant for Exploratory Research (SGER) because it represents work on an untested and novel idea that is needed urgently.

摘要玉米基因组被认为是由高度甲基化的重复DNA海洋中的轻度甲基化基因组成。由于重复的DNA，整个玉米基因组的完全测序和组装目前是不可行的。一些方法已经开始被开发出来，这些方法将允许过滤出含有这些基因的玉米基因组区域，以便进行测序。一旦在玉米中得到验证，该方法有望应用于其他复杂基因组作物的类似测序项目。初步数据表明，这种“ORF Rescue”载体系统是在复杂作物基因组中发现基因的潜在有力工具。将进行实验以验证该载体系统用于制备用于基因组测序的过滤模板DNA的效用。该项目的三个具体目标是：1）评估ORF Rescue载体从玉米基因组DNA中筛选基因的能力; 2）评估ORF Rescue载体从克隆到细菌中的玉米DNA中筛选基因的能力。人工染色体（BAC）; 3]确定新的ORF是否定义单子叶植物特异性基因。这项工作值得作为探索性研究小额赠款（SGER）提供资金，因为它代表了一个迫切需要的未经测试的新颖想法。