This project will focus on the structural characterization and generation of a physical map of human chromosome 11 as a Specialized Human Genome Research Center (P50). The five-year goal of the Center is to complete a physical map of DNA markers on chromosome 11 spaced at an average distance of 300 kb and individually identified by DNA sequence identifiers (STS """"""""sequence-tagged sites""""""""). The eventual long term goal of the Center is to complete an overlapping set of cosmid and yeast artificial chromosome (YAC) clones spanning the entire chromosome 11 which may serve as the source of template material for DNA sequencing. This project represents a close collaboration between four highly interactive research groups who will focus on cloning and mapping the chromosome 11 long arm (Evans - Project #1), short arm (Sukumar - Project #2), production of yeast artificial chromosome libraries (Hoekstra - Project #3), and the generation of somatic cell lines and techniques for fine structure regional mapping (Wahl - Project #4). The overall strategy will be to produce chromosome specific YAC and cosmid libraries, to determine the regional localization of 500 of these clones by high resolution in situ hybridization to metaphase chromosomes and analysis of various types of somatic cell hybrids, to generate identifying STS sequences and primers for analysis using the polymerase chain reaction (PCR), and to isolating bridging YAC clones from genomic libraries by multiplexed PCR. Closure of the map will be achieved by """"""""walking"""""""" one or more steps from each YAC clone. As this project will be heavily dependent on large scale repetitive procedures, five core laboratories are central to the operation of the Center and will provide technical services to all of the projects. These will include a microscopy core (Core B) for in situ hybridization mapping, a FACS core for chromosome purification (Core C); a clone analysis/archive core (Core D) for YAC library screening, long term clone storage and distribution of reagents; a DNA sequencing/synthesis core (Core E) for the determination of STS sequence information and the synthesis of PCR primers; a database core (Core F) which will collect, analyze and store data from all projects; and an administrative core (Core A) which will be responsible for the overall operation of the Center. The developmental component of the Center will support two pilot studies per year to encourage investigations in new technologies, and will recruit into the Genome Center two or three new investigators at the junior faculty level over the five-year project period. The outreach component of the Center will support 1) extensive interactions and collaborations with other investigators focusing on the physical and genetic mapping of chromosome 11 including the exchange of information and reagents, 2) interactions with other investigators involved in genome research by making accessible the core laboratory facilities, and 3) national and international focus for collaborations and interactions by hosting visiting scientists in the Genome Center Research Laboratory.
