Human-ovine comparative sequencing of a 250-kb imprinted domain encompassing the callipyge (clpg) locus and identification of six imprinted transcripts: DLK1, DAT, GTL2, PEG11, antiPEG11, and MEG8

Two ovine BAC clones and a connecting long-range PCR product, jointly spann ing similar to 250 kb and representing most of the MULGES-OY3 marker interv al known to contain the ck locus, were completely sequenced. The resulting genomic sequence was aligned with its human ortholog and extensively annota ted. Six transcripts, four of which were novel, were predicted to originate from within the analyzed region and their existence confirmed experimental ly: DLK1, DAT, GTL2, PEG11, antiPEG11, and MEG8. RT-PCR experiments perform ed on a range of tissues sampled from an 8-wk-old animal demonstrated the p referential expression of all six transcripts in skeletal muscle, which sug gests that they are under control of common regulatory elements. The six tr anscripts were also shown to be subject to parental imprinting: DLK1, DAT, and PEG11 were shown to be paternally expressed and GTL2, antiPEG11, and ME G8 to be maternally expressed.