P1 and cosmid clones define the organization of 280 kb of the mouse H-2 complex containing the Cps-1 and Hsp70 loci

Abstract

A 280 kilobase (kb) contig was isolated from mouse genomic P1 and cosmid libraries, using as probes human cDNA and genomic DNA fragments that map in the interval between the second component of complement and tumor necrosis factor genes of the HLA complex. The clone contig demonstrates synteny of eleven mouse genes that are homologous to genes initially mapped within the human major histocompatibility complex. These include the mouse homologs of BAT2 (HLA-B-associated transcript 2) through BAT9 and also three HSP70-related genes. Five P1 clones form a contig of 240 kb that spans from BAT9 through BAT3. Twelve cosmid clones are arranged in three contigs that confirm most of the structure of the P1 contig and link the mouse BAT3 homolog to the BAT2 homolog approximately 15 kb farther telomeric. Polymorphic DNA markers within the cloned region were used to map the cleft palate susceptibility-1 (Cps-1) locus to the interval between Hsp70.1 and BAT6 (valyl-tRNA synthetase). This refines the location of the Cps-1 locus to a 45 kb region contained in the H2-124 P1 insert.

?Both man & bird & beast?: comparative organization of MHC genes

“…The rest of the class II region generally follows the human pattern, except for the fact that there are two Mb genes, one of which is polymorphic, the other not ). The class III region is highly similar to the much studied human equivalent (Gasser et al 1994).…”

“…6; Grahovac et al 1992). This is in contrast to the DR and C4-CYP21 regions that show evolutionary instability (Kawaguchi et al 1991;Gasser et al 1994). Within a species, humans for example, these regions are known to exhibit some expansion and contraction.…”

“…The H2 class II and class III regions contain a similar number of genes as the equivalent human regions but are smaller in places (Gasser et al 1994). This is due to both smaller intron length as well as smaller intergenic regions.…”

Preparation and screening of an arrayed human genomic library generated with the P1 cloning system.

“…( (iii) Three hundred-500-kb P1 contigs have been generated with P1 mouse and rat libraries (11,12) that could not be generated using multiple cosmid libraries. Based on these results we believe the P1 library described here may be especially useful for cloning and mapping efforts.…”

“…former include an ability to clone DNA fragments that are up to 95 kb in size in an Escherichia coli host, an ability easily to use those clones to isolate and manipulate microgram quantities of recombinant DNA, an ability to produce libraries with a minimal effort (compared to YACs), and an ability to produce libraries that faithfully represent the genomic source DNA (9)(10)(11)(12). In addition, the P1 cloning vector, pADl0sacBII, contains a positive selection for cloning and the vector is organized such that cloned inserts are flanked by rare-cutting restriction enzymes as well as by T7 and Sp6 promoters (4).…”