A Radiation Hybrid System for the Genetic and Physical Mapping of the Corn Genome

玉米庞大而复杂的基因组阻碍了综合测绘和测序工作。然而，所有玉米基因的预期分离表明迫切需要有效的作图系统和/或降低基因组的复杂性。这项研究通过开发多功能遗传物质来满足这一需求，其中包括将DNA序列快速物理地映射到染色体和1 Mbp或更少的染色体片段的能力。这些材料与微阵列技术相结合，将允许在一组实验中绘制数千个序列的图谱。回收了全套燕麦-玉米染色体附加系。10个添加的序列与DNA的杂交允许将序列放置到染色体上，包括重复序列或多基因家族。辐射杂交(RH)系然后通过加入线的伽马辐照而产生。这项研究将专注于为所有10种附加植物获得可育系，并为所有10条染色体创建RH图谱面板。初步数据表明，将基因(EST和已知基因)有效地物理定位到染色体和染色体片段，以及识别和定位重复序列是可行的。分离在斯坦福大学的EST最初将被绘制成地图。RH物理作图将在亚染色体片段水平上为其他玉米物理作图系统提供一个支架，从而补充其他玉米物理作图系统。此次更新的具体目标是：(1)恢复一套完整的可育燕麦-玉米附加系，并对附加系和RH系的表型、细胞学、染色体稳定性和遗传特性进行鉴定；(2)为所有10条单独的染色体制作RH图谱，用于低分辨率和高分辨率作图；(3)使EST高通量定位到染色体区域，用于候选基因发现和比较基因组研究。交付成果：1.每个玉米染色体的可育燕麦-玉米附加系；用于分配的附加系的种子、组织或DNA。2.辐射杂交系的DNA板，用于每条染色体的低分辨率和高分辨率作图。每条染色体的辐射杂交作图面板，以及适当的资源，使其他研究人员能够将基因物理作图到1到5兆基以内。一个定位到染色体和辐射杂交系的序列数据库，当这些系可用时。要提供给现有网络资源的数据。利用来自燕麦-玉米附加系的基因组DNA，利用微阵列技术测量每条染色体重复序列的相对数量的数据。描述遗传物质、辐射杂交图谱和序列图谱的网络资源。项目地址：明尼苏达大学，系。农学和植物遗传学411Borlag Hall，1991年上Buford Circle，St.Paul，MN 55108项目信息网址(网页和电子邮件)：http://agronomy.coafes.umn.edu/cornpep/nsf/

The large and complex genome of corn has impeded integrated mapping and sequencing efforts. The anticipated isolation of all the corn genes, however, indicates the pressing need for efficient mapping systems and/or a reduction in genome complexity. This research addresses the need by developing multifunctional genetic materials that include the ability to rapidly physically map DNA sequences to chromosomes and chromosome segments of 1 Mbp or less. These materials coupled with microarray technology will allow the mapping of thousands of sequences in a single set of experiments. The complete set of oat-corn chromosome addition lines has been recovered. Hybridization of sequences to DNA of the 10 additions allows placement of sequences to chromosome including duplicated sequences or multigene families. Radiation hybrid (RH) lines are then produced by gamma irradiation of the addition lines. The research will focus on obtaining fertile lines for all 10 addition plants and on creating RH mapping panels for all 10 chromosomes. Preliminary data indicates the feasibility for efficient physical mapping of genes (ESTs and known genes) to chromosome and chromosome segment, and the identification and mapping of duplicated sequences. The ESTs isolated at Stanford University will initially be mapped. An RH physical map will complement other corn physical mapping systems by providing a scaffold at the subchromosomal fragment level. The specific objectives of this renewal are to (1) recover a complete set of fertile oat-corn addition lines and characterize the phenotype, cytology, chromosome stability and transmission of the addition and RH lines; (2) produce RH mapping panels for all 10 individual chromosomes for both low and high resolution mapping; (3) enable high-throughput mapping of ESTs to chromosomal regions for candidate gene discovery and comparative genome studies.Deliverables:1. Fertile oat-corn addition lines for each corn chromosome; seed, tissue or DNA of the addition lines for distribution. 2. DNA panels of radiation hybrid lines for low and high-resolution mapping of each chromosome.3. Radiation hybrid mapping panels for each chromosome, along with appropriate resources that will allow other researchers to physically map genes to within one to five megabases.4. A database of sequences mapped to chromosome and to radiation hybrid lines as the lines become available. Data to be provided to existing web resources.5. Data about relative quantities of repetitive sequences per chromosome measured using microarray technology with genomic DNA from the oat-corn addition lines.6. Web resources describing genetic materials, radiation hybrid mapping, and sequences mapped.Project address: University of Minnesota, Dept. of Agronomy and Plant Genetics 411 Borlaug Hall, 1991 Upper Buford Circle, St. Paul, MN 55108 Internet Addresses (Web and E-mail) for Project Information: http://agronomy.coafes.umn.edu/cornpep/nsf/