Figure 7From: Revised genomic structure of the human ghrelin gene and identification of novel exons, alternative splice variants and natural antisense transcriptsCharacterisation of ghrelin natural antisense transcripts in the normal human stomach. A. Ethidium bromide stained agarose gel showing the verification of the candidate antisense gene ghrelinOS by orientation-specific RT-PCR in the human stomach. Antisense transcripts were amplified using a primer in exon 4* as the reverse transcription (RT) primer (R), while for the detection of sense transcripts, a primer in exon 2* was used in the RT reaction (F). The RT reactions were subjected to PCR using both the exon 2* and 4* primers (for 30, 35 or 40 cycles). NTC = no template control (water). M = MassRuler Express DNA ladder (Fermentas). B. Alignment of sequences derived from two 5' RLM-RACE products (TSS86 and TSS63) and a CAGE tag starting site corresponding to a 28 bp exon 4* (TSS28, T03F009D342E) with exon 4* and flanking genomic sequence. The lower case letters indicate upstream genomic or downstream intron 4* sequences. The position and sequence of the nested gene-specific 5' RACE primer (5'OS-in-R) is indicated with an arrow and underlined, respectively. For comparison, the (sense) ghrelin gene exon 4 sequence is shaded in grey.Back to article page