The purpose of this project is to utilize a combination of somatic cell genetic and recombinant DNA technologies to carry out fine structure gene mapping of two human chromosomes, numbers 5 and 12. These two chromosomes were chosen for study because genetic selective systems are available which make possible fine structure mapping of them using recombinant DNA probes. For each of these chromosomes there is available: a) a CHO-human hybrid cell line which retains, under selective pressure, the single human chromosome under study; b) a complete recombinant DNA library specific for each of the two chromosomes, and c) a selective system to isolate a large number of derivatives of the single human chromosome-containing hybrid which have deletions of different, well-defined parts of the chromosome. Low copy DNA fragments from the human chromosome-specific recombinant libraries will be identified and subcloned and will be used as probes for Southern blots of restriction endonuclease digested DNAs from the series of cell lines with deletions of various parts of the chromosome under study. One can then correlate the presence or absence of human specific DNA fragments that hybridize to the various probes tested with the regions of the chromosome that are deleted in the various cell lines. This approach to mapping the human genome provides an opportunity to construct very detailed fine structure maps of human chromosomes. DNA probes localized to specific regions if one of these two chromosomes will then be screened to identify those that detect common restriction fragment length polymorphisms (RFLP) in members of a large human kindred. The goal of this part of the project is to use common RFLP as genetic markets to examine, for a number of pairs of loci, the relationship between positions of, and physical distances between, genes to meiotic recombination frequencies. Studies on recombination for a number of different regions along a given chromosome may well define regions in which recombination is much more, or much less frequent than in other regions, which could potentially help lead to an understanding of the structural or sequence elements of DNA that affect recombination. A related part of the project is aimed at using three other recombinant DNA libraries, specific for human chromosomes, 3, 4, and 18, respectively, to identify DNA probes from each chromosome that detect common RFLP in humans, which will be very useful for examining linkage relationships in families with genetic disorders in which the chromosomal location of the altered gene is unknown.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM025339-07
Application #
3272945
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1978-08-01
Project End
1989-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
Schools of Medicine
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697
Farber, R A; Phalen, T; Neuman, W L et al. (1988) An anonymous DNA segment pTP5E (D5S70) maps to the long arm of chromosome 5 and identifies a Taq I polymorphism. Nucleic Acids Res 16:2360
Smith, B; Skarecky, D; Bengtsson, U et al. (1988) Isolation of DNA markers in the direction of the Huntington disease gene from the G8 locus. Am J Hum Genet 42:335-44
Dobbs, M; Overhauser, J; Wasmuth, J J (1988) Molecular analysis of a case of meiotic recombination leading to cri-du-chat syndrome. Cytogenet Cell Genet 47:5-7
Carlock, L R; Smith, D; Wasmuth, J J (1986) Genetic counterselective procedure to isolate interspecific cell hybrids containing single human chromosomes: construction of cell hybrids and recombinant DNA libraries specific for human chromosomes 3 and 4. Somat Cell Mol Genet 12:163-74
Nakamichi, N N; Kao, F T; Wasmuth, J et al. (1986) Ribosomal protein gene sequences map to human chromosomes 5, 8, and 17. Somat Cell Mol Genet 12:225-36
Wasmuth, J J; Carlock, L R; Smith, B et al. (1986) A cell hybrid and recombinant DNA library that facilitate identification of polymorphic loci in the vicinity of the Huntington disease gene. Am J Hum Genet 39:397-403
Wasmuth, J J; Carlock, L R (1986) Chromosomal localization of human gene for histidyl-tRNA synthetase: clustering of genes encoding aminoacyl-tRNA synthetases on human chromosome 5. Somat Cell Mol Genet 12:513-7
Mohandas, T; Heinzmann, C; Sparkes, R S et al. (1986) Assignment of human 3-hydroxy-3-methylglutaryl coenzyme A reductase gene to q13----q23 region of chromosome 5. Somat Cell Mol Genet 12:89-94
Maurer, B J; Carlock, L; Wasmuth, J et al. (1985) Assignment of human dihydrofolate reductase gene to band q23 of chromosome 5 and of related pseudogene psi HD1 to chromosome 3. Somat Cell Mol Genet 11:79-85
Carlock, L R; Skarecky, D; Dana, S L et al. (1985) Deletion mapping of human chromosome 5 using chromosome-specific DNA probes. Am J Hum Genet 37:839-52