International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)

A4. Genomic Sequence Analysis in the Mouse T-complex Region

Ramaiah Nagaraja, Paul Waeltz, Eudora Jones, Suzelle M. Amyot, Yong Qian, Dawood Dudekula, Mgavi Brathwaite, Minoru S.H. Ko and David Schlessinger.
Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD 21224, USA

A systematic analysis of 30 Mb of the mouse t-complex region on chromosome 17 has been undertaken to define the regional content of genes, including candidates for t-locus mutations, and to help determine their role in early embryonic development. Physical map construction was initiated using ESTs from a collection derived from preimplantation (Development 127, 1737-1749, 2000) and 7.5 dpc embryos (Human Molecular Genetics 7, 1967-1978, 1998), along with STSs designed from other gene sequences mapped to this region. These were used as probes to screen the RPCI-23 mouse BAC library. Additional STSs generated from end sequences of BACs and from interB1-products confirmed the integrity of recovered clones and the extent of their overlap in the nucleated contigs. Reiterated screenings from the BAC-end STSs provide clones from adjacent regions and extend the contigs. BACs contigs now cover 2.2 Mb of the region with 6-fold multiplicity. Growing contigs are being extended and will be further integrated with overlapping mouse genomic sequencing that may come from other sites.

To date, seven BACs have been sequenced, generating 1.2 Mb of nonoverlapping sequence. Details of the genomic and gene structure for sequenced clones will be presented. The sequence and complete analysis of one BAC, 126c8, has been recently published (Genome Research, 10, 916-923, 2000). Salient features of the other six included the placement of known but previously unlocalized genes Clc7, Zfp118 (KRAZ1), and Pdpk1 on chromosome 17. Furthermore, several clusters of gene family members have been analyzed. One includes the zinc finger protein gene Zfp118 along with zinc finger protein genes ZFP 54 and ZFP51. A second includes 3 solute carrier proteins (Slc22a1, Slc22a2, Slc22a3) between Plg and Igf2r.

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