Figure 4.

Expression of an antisense transcript associated with KvDMR1. (a) RNA was isolated from human fetal tissues and analyzed by RT-PCR with the indicated primer pairs (see Fig. 2a for location of primers). Primers 686.3 and 592.20 were designed from EST 68627 and EST 592241, respectively, and were used to show the connection of these two ESTs. The + and − indicate that the PCR templates were from cDNA (+RT) or mock cDNA (−RT), respectively. (b, Upper) Southern blot of BamHI/NotI-digested DNA from a panel of eight single human chromosome 11 somatic cell hybrids (47) hybridized with DMRP. (b, Lower) RT-PCR analysis of same hybrids with primers specific for EST 592241. (c) Paternal-specific expression at the mouse KvDMR1 locus. Arrows indicate the presence of both alleles in DNA from F₁ animals. In F₁ fetal liver RNA from a C57BL/6J × PWK mating, as well as in the reciprocal cross, only the paternal allele was detected. The weak bands in the −RT lane result from contaminating DNA in the RNA samples, because the PCR primers do not amplify across an intron.