SGER: An ORF - Rescue Vector for Crop Genome Sequencing

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的岛。由于DNA的重复，玉米全基因组的完整测序和组装目前是不可行的。正在开始开发方法，允许筛选出玉米基因组中包含基因的区域进行测序。这种方法一旦在玉米中得到验证，有望用于其他具有复杂基因组的作物物种的类似测序项目。一种新的表达载体系统已经开发出来，可以直接从基因组DNA中回收开放阅读框架(ORF)。初步数据表明，这种“ORF Rescue”载体系统是发现复杂作物基因组中基因的一种潜在的强大工具。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的实用性。该项目的三个具体目标是：1)评估ORF救援载体从玉米基因组DNA中筛选基因的能力；2)评估ORF救援载体从克隆到细菌人工染色体(BAC)中的玉米DNA中筛选基因的能力；3)确定新的ORF是否定义了单子叶植物特有的基因。这项工作值得作为探索性研究小额赠款(SGER)提供资金，因为它代表了一种迫切需要的未经检验的新想法的工作。

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.