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 spanning approximately 250 kb and representing most of the MULGE5-OY3 marker interval known to contain the clpg locus, were completely sequenced. The resulting genomic sequence was aligned with its human ortholog and extensively annotated. Six transcripts, four of which were novel, were predicted to originate from within the analyzed region and their existence confirmed experimentally: DLK1, DAT, GTL2, PEG11, antiPEG11, and MEG8. RT-PCR experiments performed on a range of tissues sampled from an 8-wk-old animal demonstrated the preferential expression of all six transcripts in skeletal muscle, which suggests that they are under control of common regulatory elements. The six transcripts were also shown to be subject to parental imprinting: DLK1, DAT, and PEG11 were shown to be paternally expressed and GTL2, antiPEG11, and MEG8 to be maternally expressed.
Charlier
C
C
Department of Genetics, Faculty of Veterinary Medicine, University of Liège (B43), 4000-Liège, Belgium.
Segers
K
K
Wagenaar
D
D
Karim
L
L
Berghmans
S
S
Jaillon
O
O
Shay
T
T
Weissenbach
J
J
Cockett
N
N
Gyapay
G
G
Georges
M
M
D000818:Animals; D001482:Base Composition; D019295:Computational Biology / Q000379:methods; D018899:CpG Islands / Q000235:genetics; D003596:Cytosine / Q000032:analysis; D020224:Expressed Sequence Tags; D005819:Genetic Markers; D018392:Genomic Imprinting / Q000235:genetics*; D006147:Guanine / Q000032:analysis; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000502:physiology; D011506:Proteins / Q000235:genetics; D062085:RNA, Long Noncoding; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep / Q000235:genetics*; D014158:Transcription, Genetic
D005819:Genetic Markers; D047908:Intracellular Signaling Peptides and Proteins; C498907:MEG3 non-coding RNA, human; D008565:Membrane Proteins; D011506:Proteins; D062085:RNA, Long Noncoding; C064045:delta protein; D006147:Guanine; D003596:Cytosine