Figure 3

Verification of an extended exon 1 in the mouse ghrelin gene. A. Schematic diagram showing the structure of the murine ghrelin gene. Exons are represented as boxes and introns as horizontal lines. PCR primers employed in B. and C. are shown as arrows and the exon regions amplified are indicated. B. Ethidium bromide stained agarose gel electrophoresis demonstrating Ghrl extended exon 1 to exon 1 RT-PCR amplicons. DNase treated total RNA was amplified by RT-PCR with reverse transcriptase (+RT), or without (-RT) reverse transcriptase to demonstrate that there was no genomic DNA contamination. Gapdh was used as an endogenous control. C. Ethidium bromide stained agarose gel electrophoresis of a 445 bp full-length preproghrelin amplicon derived using primers located in the extended exon 1 (Ext1-F) and exon 4 (Ex4-R). M = 100 bp DNA molecular weight marker ladder (New England Biolabs). NTC = no template negative control.