International Mammalian Genome Society


The 13th International Mouse Genome Conference
October 31-November 3, 1999

Table of Contents * Structure * Bioinformatics * Sequence * Mapping * New Tools * Gene Discovery * Developmental * Mutagenesis * Functional Genomics

D21 Analysis of Regulatory Elements for Pax1 and Pax9 by BAC Transgenesis and Comparative Sequencing

Imai, K.1, Kokubu, C.1, Santagati, F.1, Wilm, B.1, Peters, H.1, Dietz, U.1, Gerber, J.-K.1, Suzuki, M.2, Abe, K.2, Yamamura, K.2, Balling, R.1 1GSF-National Research Center for Environment and Health, Institute of Mammalian Genetics, Ingolstaedter Landstr. 1, 85764 Neuherberg, Germany; 2Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine, 4-24-1 Kuhonji, Kumamoto 862-0976, Japan

Pax1 and Pax9 constitute a subfamily of the Pax gene family, characterized by the paired-box, a sequence coding for a motif with DNA binding activity. Pax1 and Pax9 are not only highly related at the sequence level, but also share expression domains within the developing sclerotome and the pharyngeal pouch endoderm. Therefore it is reasonable to assume that similar cis-acting control elements might direct expression of Pax1 and Pax9. Here we report our efforts towards identification of regulatory elements for Pax1 and Pax9 by two parallel approaches: BAC transgenesis and comparative sequencing. Our classical transgenic approach with a 14 kb fragment upstream of the first exon for Pax9 failed to reproduce endogenous expression patterns. We also could not rescue the Pax1-deficient phenotype by transgenesis with Pax1-BACs (75-135 kb) including 25-100 kb upstream regions of Pax1. These observations led us assume that genome units including all exons, promoter and regulatory elements for Pax1 and Pax9 might be very large. Therefore we decided to obtain BACs with larger inserts to extend our BAC transgenic approach. Eight and twelve BACs have been isolated for Pax1 and Pax9, respectively. As a first step, we have established long-range genome maps for the Pax1 and Pax9 loci with these BACs. In parallel, sequencing mouse Pax1 and Pax9 genomic regions is in progress. By database search, we could find large genome sequences including human PAX1 or PAX9. We will present data from sequence comparison between human and mouse Pax1/9.

 


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