Genomic structure of chick IRK1 and determination of transcription initiation sites (A) Restriction map of chick IRK1/Kir2.1 genomic locus. The four overlapping genomic clones are shown above the restriction map. Solid boxes indicate exons. E; EcoRI site, X; XbaI site. The probe used for northern blots is underlined. (B) Genomic sequence surrounding splice sites. Exon 1 ends at +103 and exon 2 begins at +104 (based on cIRK1 cDNA, U20216), as indicated. Intronic sequences are in italics; bold letters indicate donor site GT and acceptor site AG. (C) Primer extension analysis. Approximate size, in bases, is indicated on the left. Products of about 80 bases were present in brain, but not in DF1 fibroblasts or in controls (tRNA). (D) Determination of transcription initiation sites using 5'RACE. Shown are the results of the secondary PCR using the 5'-nested primer and a primer designed at 64-41 of cIRK1 cDNA (N-R2), and controls using the 5'-nested primer only (N only) or the reverse primer only (R2 only).
Mutai et al. BMC Genomics 2004 5:90 doi:10.1186/1471-2164-5-90