The overall objective of this program project is to apply newly emerging technologies to the large scale physical and genetic mapping of the human genome and the generation of ordered sets of cloned DNA spanning megabase regions of the X chromosome. Specific objectives are: 1)to generate a physical linkage map of the human genome (all chromosomes) by precisely localizing about 6000 cloned DNA or cDNA fragments on metaphase chromosomes by multiparameter in situ hybridization using fluorescence digital imaging microscopy. These DNAs will include about 5000 random phage and cosmid clones, 500 clones known to contain a RFLP, 300 Cla/Cla linking clones (with spacing of 5-10 mega bp) as well as several hundred additional Not/Not or Taq/Taq linking library clones. 2)to carry out genetic linkage studies on 100 DNA markers per year by hybridization to CEPH and other large reference family pedigrees and to screen about 200 clones per year from the panel of clones mapped by in situ hybridization for RFLPs and a subset of these for other types of sequence polymorphism. 3)to prepare a complete set of genomic Cla-Cla linking fragments and a complete set of Taq-Taq or BsuE-methylated Not I for the X chromosome. 4)to apply affinity-""""""""fishing""""""""-techniques to the direct isolation of ordered DNA fragments from the X chromosome and to prepare clone sets for each fragment. 5)to prepare a cDNA expression map of the X chromosome using a) in situ hybridization with cDNA clones, b) cross-library screening to identify tissue-specific patterns of expression (cDNA libraries from multiple human tissues have already been prepared) and c) cDNA library screening with selected clones prepared from affinity purified Cla/Cla DNA fragments. 6)to sequence the panel of cDNA clones identified and mapped as part of research objective #5._

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
3R01HG000272-03S1
Application #
3333340
Study Section
Special Emphasis Panel (SRC)
Project Start
1990-05-01
Project End
1993-12-02
Budget Start
1993-05-01
Budget End
1993-12-02
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Henegariu, O; Dunai, J; Chen, X N et al. (2001) A triple color FISH technique for mouse chromosome identification. Mamm Genome 12:462-5
Cremona, O; Nimmakayalu, M; Haffner, C et al. (2000) Assignment of SYNJ1 to human chromosome 21q22.2 and Synj12 to the murine homologous region on chromosome 16C3-4 by in situ hybridization. Cytogenet Cell Genet 88:89-90
Wang, H; Chatterjee, G; Meyer, J J et al. (1999) Characteristics of three homologous 202 genes (Ifi202a, Ifi202b, and Ifi202c) from the murine interferon-activatable gene 200 cluster. Genomics 60:281-94
Lizardi, P M; Huang, X; Zhu, Z et al. (1998) Mutation detection and single-molecule counting using isothermal rolling-circle amplification. Nat Genet 19:225-32
Jacob, A N; Manjunath, N A; Bray-Ward, P et al. (1998) Molecular cloning of a zinc finger gene eZNF from a human inner ear cDNA library, and in situ expression pattern of its mouse homologue in mouse inner ear. Somat Cell Mol Genet 24:121-9
Kovalenko, O V; Golub, E I; Bray-Ward, P et al. (1997) A novel nucleic acid-binding protein that interacts with human rad51 recombinase. Nucleic Acids Res 25:4946-53
Haaf, T; Ward, D C (1996) Inhibition of RNA polymerase II transcription causes chromatin decondensation, loss of nucleolar structure, and dispersion of chromosomal domains. Exp Cell Res 224:163-73
Marondel, I; Renault, B; Lieman, J et al. (1996) Physical mapping of the human neurotensin gene (NTS) between markers D12S1444 and D12S81 on chromosome 12q21. Genomics 38:243-5
Cupelli, L; Renault, B; Leblanc-Straceski, J et al. (1996) Assignment of the human myogenic factors 5 and 6 (MYF5, MYF6) gene cluster to 12q21 by in situ hybridization and physical mapping of the locus between D12S350 and D12S106. Cytogenet Cell Genet 72:250-1
Li, M J; Peakman, M C; Golub, E I et al. (1996) Rad51 expression and localization in B cells carrying out class switch recombination. Proc Natl Acad Sci U S A 93:10222-7

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