Abstracts: 

 Physical Mapping 



In collaboration with D. Ward and P. Lichter (Yale University Medical School) and 

 D. Housman and K. Call (Massachusetts Institute of Technology), we have used high- 

 resolution in situ hybridization of single cosmid clones to map selected contigs and 

 landmark clones to chromosomal locations. Produced in the pilot study, the resulting 

 collection of clones, containing over 609f of the 1 lql2 to I Iqter region, is contained in 

 a reference collection now undergoing analysis. To expand this pilot study to a larger 

 collection of clones representing a tenfold redundancy of the entire chromosome 1 1 , we 

 have used a fluorescence-activated cell sorter to purify human chromosome 1 1 from a 

 somatic cell hybrid, Jl, containing a single chromosome 1 1 in a CHO-Kl cell 

 background, and we are preparing a chromosome-1 1 -specific cosmid library in sCos-1. 



This work represents the beginning of a large-scale mapping project to obtain, 

 reference, archive, and link cosmids spanning the entirety of human chromosome 11, a 

 project which may be complemented by studies using pulsed-field gel electrophoresis 

 and yeast artificial chromosomes. During the next year, the Center for Human Genome 

 Research will expand its program to include additional Salk Institute investigators 

 interested in gene recombination and amplification. YAC vectors and mapping 

 strategies, novel in vivo mapping techniques, and identification of recessive oncogenes. 

 The goals are to complete the chromosome-1 1 physical map and proceed with 

 characterization of genes important to human biology. New methodologies to be 

 utilized in the longer temi include DNA sequencing, functional expression in 

 mammalian cells in culture, creation of transgenic mouse strains as models for human 

 disease states, and the identification of functional genes by genetic complementation. 



The immediate goals are to ( 1 ) improve engineering, robotic, and computational 

 systems for preparation and management of arrayed cosmids and their subsequent 

 processing: (2) expand the arrayed library of chromosome-1 1 cosmids to 

 10,000-33.000 members: (3) establish a corresponding repository and database for the 

 distribution of these resources and the correlation of results from other laboratories: 

 (4) pursue detection and characterization of expressed genes, especially those relevant 

 to disease, while completing the physical map: (5) continue the conceptual and practical 

 development of the multiplex strategy including extension of the probe-pooling strategy 

 from two- to higher-dimensional arrays of the cosmids, establishment of the optimal 

 library size, further suppression of effects of troublesome high-copy-number sequences, 

 and exploration of the utility of PCR techniques for probe preparation: (6) develop a 

 correlative approach to integration of chromosome-1 1 map data acquired through 

 multiplex walking, PFG and linking clone analyses, linkage data obtained through the 

 use of a variable number of tandem repeats ( VNTR), RFLP, and minisatellite probes 

 and radiation hybrids: and (7) continue the characterization of DNA/genes adjacent to 

 the chromosome-1 1 translocation breakpoints obtained from clinical sources and pursue 

 the identification of disease genes mapped to chromosome 1 1. 



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