Figure 4
From: Unusual Phenotypic Features in a Patient with a Novel Splice Mutation in the GHRHR Gene
Transcription assay. (A) RT-PCR amplification of the GHRHR transcripts obtained from cells transfected with plasmids pcDNA3.1-GHRHR_WT (wild type minigene, amplified between P1-P2) (Lane wt) and pcDNA3.1-GHRHR_mut (mutated minigene, amplified between P1–P2) (Lane mut) using two internal exonic primers (P3 and P4), indicated by arrows in B (a, c). (B), (b) Schematic representation of the genomic GHRHR region where the IVS1 + 2T > G mutation at the donor splice site (gtgagta > gggagta) and the cryptic 5′ splice site (gtaag) (428bp downstream the first nucleotide of intron 1) are in bold characters. The 290-bp product corresponds to the normal transcript and the 718-bp fragment to the mutated transcript with a retention of a part of intron 1 because of activation of the intronic cryptic donor splice site (bold characters). (a) and (c): schematic representation of mutated and wild type partial cDNAs, respectively.