MAMMALIAN GENOME MAPPING OF TISSUE-SPECIFIC CDNAS

Project: Research project

Description

Tissue specific cDNAs will be rapidly mapped to mouse chromosomal segments
in order to facilitate the definition of both mouse and human genomic
organization. An interspecific mouse cross between Mus spretus and a
laboratory inbred strain will allow definition of cDNA RFLV markers and
mapping by haplotype analysis of most if not all cDNA clones. Equalized
neonatal thymus specific cDNA libraries will be obtained by the
application of a novel solid phase system in which cDNAs will be bound and
eluted from immobilized genomic sequences. This technique should allow the
generation of a library of tissue specific cDNAs that cross hybridize
between mammalian species by the selection mouse cDNAs that bind to human
genomic sequences. Unique cDNAs (approximately 1000) will then be mapped
after PCR amplification of sequences. A panel of 38 interspecific
backcross mouse DNAs that has been characterized for over 45O markers
throughout the mouse genome will be used for rapid mapping. This panel
divides the mouse genome into over 470 different segments. A stratified
mapping approach will be subsequently utilized to divide the genome into
1000 different segments. In addition, reference markers at 5 cM intervals
that have been analyzed in 100 to 400 meiotic events will further define
the mapped location of new cDNA markers. The results will provide a
putative human regional localization by the application of comparative
mapping knowledge. Selected cDNAs, a total of 200, will be used to
precisely define the borders of conserved linkage groups by in situ
hybridization on human chromosomes. The cDNA clones will also be partially
sequenced to provide sequenced tagged sites, allow PCR screening of YAC
libraries, and enable the mapping of single stranded conformational
polymorphisms by interested investigators. Since 5' as well as 3' sequence
information will be obtained, these transcripts will be compared with
previously characterized genes. Novel transcripts will provide a resource
for molecular genetic studies of thymocyte maturation. In addition, the
mapping of over 1000 unique neonatal thymus transcripts may determine
whether there are regions of the genome that are transcriptionally active
in a coordinate fashion. The mapped cDNA clones will be made easily
available to other investigators and should dramatically facilitate
closure of long range restriction maps as well as provide additional tools
for contig construction of gene rich regions of both the mouse and human
genome. The cDNA map will also be integrated with the developing
microsatellite STS maps in other laboratories. This will allow integration
of efforts to define a contig of the mouse genome.
StatusFinished
Effective start/end date9/5/938/31/00

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $374,459.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

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Chromosome Mapping
Complementary DNA
Genome
Expressed Sequence Tags
Genes
Clone Cells
Research Personnel
Thymus Gland
Microsatellite Repeats
Databases
Polymerase Chain Reaction

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)