Molecular Cloning of H-2 Class I Genes in the H-2b Haplotype
We have cloned H-2 class I genes of the C57BL/10 (B10, H-2b haplotype) mouse using cosmid cloning techniques developed in our laboratory (1). Cosmids, which contain 40–45 kilobasepairs (kbp) of mouse DNA per clone, were chosen since there are about 20–30 class I genes (see below) in the mouse genome and most of these genes map between the H-2K and TL loci on chromosome 17 (Figure 1); a region of some 1000 kbp. of DNA. Estimates of the number of class I genes in the mouse genome are based on Southern blot analysis (2) in which restriction enzyme digested mouse genomic DNA is hybridised to a class I H-2 cDNA probe (Figure 2). In all digests shown, and in all three H-2 haplotypes tested, the cDNA probe hybridises to multiple bands in the digests indicating that the H-2 class I are multiple copy and highly homologous. Using either H-2 cDNA or H-LA genomic probes we have isolated 90 cosmids containing class I genes from a mouse genomic library constructed using B10 spleen DNA. These cosmids have been organised into 5 distinct clusters of overlapping mouse DNA on the basis of restriction enzyme mapping and Southern blot hybridisation analysis (Table 1). The DNA regions cloned in each cosmid cluster have been mapped to one of the four genetic loci known, from immunological analysis, to control the expression of class I cell surface antigens in the B10 mouse (see Figure 1).
KeywordsCell Surface Antigen Restriction Enzyme Mapping Southern Blot Hybridisation Analysis Specific Genetic Locus Recombinant Inbred Mouse Strain
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