Genetic and physical mapping of the Lps locus: Identification of the toll-4 receptor as a candidate gene in the critical region

Alexander Poltorak, Irina Smirnova, Xiaolong He, Mu Ya Liu, Christophe Van Huffel, Dale Birdwell, Erica Alejos, Maria Silva, Xin Du, Patricia Thompson, Edward K L Chan, Jessica Ledesma, Bruce Roe, Sandra Clifton, Stefanie N. Vogel, Bruce Beutler

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Abstract

On the basis of 2093 meioses analyzed in two separate intraspecific backcrosses, the location of the mouse Lps(d) mutation was circumscribed to a genetic interval 0.9 cM in size. A total of 19 genetic markers that lie in close proximity to the mutation were examined in mapping. Most of these were previously unpublished polymorphic microsatellites, identified by fragmentation of YAC and BAC clones spanning the region of interest. Lps(d) was found to be inseparable from the microsatellite marker D4MIT178, and from three novel polymorphic microsatellites identified near D4MIT178. The mutation was confined between two novel microsatellite markers, herein designated 'B' and '83.3.' B lies centromeric to the mutation, and was separated by four crossovers in a panel of 1600 mice; 83.3 lies distal to the mutation and was separated by three crossovers in a panel of 493 mice. 66 BAC clones and one YAC clone were assembled to cover >95% of the critical region. Estimates based on pulsed field gel electrophoresis and fluorescence in situ hybridization indicate that the The B→ 83.3 interval is about 3.2 Mb in length. A minimal area of zero recombinational distance from Lps(d) was also assigned, and found to occupy approximately 1.2 Mb of physical size. To identify gene candidates, nearly 40,000 sequencing runs were performed across the critical region. Selective hybridization and exon trapping were also employed to identify genes throughout the 'zero' region. Only a single intact gene was identified within the entire critical region. This gene encodes the Toll-4 receptor, a member of the IL- 1 receptor family.

Original languageEnglish (US)
Pages (from-to)340-355
Number of pages16
JournalBlood Cells, Molecules, and Diseases
Volume24
Issue number3
DOIs
StatePublished - Sep 1998

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Keywords

  • Endotoxin
  • Inflammation
  • Lipopolysaccharide
  • Positional cloning
  • Sepsis
  • Shock

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Hematology
  • Cell Biology

Cite this

Poltorak, A., Smirnova, I., He, X., Liu, M. Y., Van Huffel, C., Birdwell, D., Alejos, E., Silva, M., Du, X., Thompson, P., Chan, E. K. L., Ledesma, J., Roe, B., Clifton, S., Vogel, S. N., & Beutler, B. (1998). Genetic and physical mapping of the Lps locus: Identification of the toll-4 receptor as a candidate gene in the critical region. Blood Cells, Molecules, and Diseases, 24(3), 340-355. https://doi.org/10.1006/bcmd.1998.0201