THE CONSTRUCTION OF HUMAN SOMATIC-CELL HYBRIDS CONTAINING PORTIONS OF THE MOUSE X-CHROMOSOME AND THEIR USE TO GENERATE DNA PROBES VIA INTERSPERSED REPETITIVE SEQUENCE POLYMERASE CHAIN-REACTION

Interspersed repetitive sequence polymerase chain reaction (IRS-PCR) has become a powerful tool for the rapid generation of DNA probes from human chromosomes present in rodent somatic cell hybrids. We have constructed a somatic cell hybrid containing a major portion of the mouse X chromosome in a human background (clone 8.0). IRS-PCR was developed for the specific amplification of mouse DNA using either of two primers from the rodent-specific portion of the murine B1 repeat. Amplification was subsequently performed with clone 8.0 and a subclone, 8.1 1, which retains a small murine X-chromosomal fragment including Hprt and the Gdx locus. A total of 15-20 discrete PCR products ranging in size from <500 to >3000 bp were obtained from clone 8.0 with each primer. In clone 8.1 1, a subset of these bands plus some additional bands were observed. Nine bands amplified from clone 8.1 1 have been excised from gels and used as probes on Southern blots. All of the fragments behaved as single-copy probes and detected domesticus/spretus variation. They have been regionally mapped using an interspecific backcross. The probe locations are compatible with those of markers known to be present in clone 8.1 1. These results demonstrate the feasibility of this method as applied to the mouse genome and the high likelihood of generating useful DNA probes from a targeted region. © 1991.