Construction of a detailed molecular map of the mouse X chromosome by microcloning and interspecific crosses.

Abstract

A large number of microclones obtained by microdissection of the mouse X chromosome have been mapped using an interspecific Mus domesticus/Mus spretus cross. Clones displaying close linkage to a number of loci of known phenotype but unknown gene product, such as mdx (X‐linked muscular dystrophy), have been obtained. Over a central 30 cM span of the mouse X chromosome, 17 clones have been mapped and ordered at a sufficient density to contemplate the complete physical mapping of this region that will aid in the isolation of a number of unidentified genes. Some of the mapped microclones detect moderately repetitive sequences that were clustered in several discrete regions of the mouse X chromosome.

Localization of the mdx mutation within the mouse dystrophin gene.

“…The order of loci was determined by analysis of individual animals with recombinational breakpoints between given loci (Figure 1). The frequencies of recombination and the order of the loci are in agreement with those from three previous studies (Amar et al, 1985;Brockdorff et al, 1987b;Heilig et al, 1987) between pD38 and mdx, indicating that the mdx mutation Partial cDNA clones corresponding to segments of the is telomeric to pD38 (Table 11). A HindJII-EcoRI fragment, human DMD transcript have also been mapped through the -1.0 kb in length, detecting the last exon at the 3' end of pedigree relative to mdx and the eight other loci.…”

“…Inter-species crosses between Mus musculus and M. spretus have recently been shown to be of value in the mapping of genes and marker DNA sequences on the mouse X chromosome (Amar et al, 1985;Avner et al, 1987;Brockdorff et al, 1987b;Chamberlain et al, 1987;Heilig et al, 1987). In such crosses the evolutionary divergence of the two genomes greatly increases the frequency of restriction fragment length polymorphisms, facilitating the detection and analysis of meiotic recombinations between loci.…”

Unusual molecular characteristics of a repeat sequence island within a Giemsa-positive band on the mouse X chromosome.

“…An interspecific Mus domesticus/Mus spretus cross segregating for the coattexture mutations Hq, Ta, and Li (13) was used to generate backcross progeny for the genetic analysis of DXSmh141 and other microclones and has been described in detail (14,15). High molecular weight DNA was prepared from mouse tails (14).…”

“…Hybridization of DXSmh141 to genomic digests of M. spretus DNA under low-stringency conditions failed to reveal further bands (data not shown). (14). Progeny were scored for the coattexture mutations and a variety of microclone and genic probes including DXSmh141.…”

The Development of Chromosome Microdissection and Microcloning Technique and its Applications in Genomic Research

“…Then, it was extended to human chromosomes [3]. However, at that time, studies were mainly focusing on some chromosomes that are easily identifiable by their configuration, such as the X chromosome of mouse [5] and chromosome 2 of human [3]. After the advent of G-banding technique, which makes the identification of human and animal chromosomes easier, and PCR technique, the chromosome microdissection and microcloning technique was extensively used in human and animal genomics research [38,46,47,49,74,75].…”