PEG10 transcript analysis reveals the existence of two splice variants.
A. Shown is the sequence around the splice junctions for the two PEG10 splice variants. A predicted ATG start codon in exon 2 is underlined. The exon 1b transcript represents a rare splice event, which leads to a new ATG start codon (underlined) in exon 1b. B. In order to estimate the frequency of PEG10 alternative splicing PEG10-specific PCR was done with cell line HepG2 and SH-SY5Y cDNA using forward and reverse primers located in exon 1 and exon 2 respectively. PCR products were cloned and transformed into bacteria. 9 colonies were analysed by colony PCR with the ABP-F2/ABP-R2 primer pair that allows to amplify both splice variants. PCR products were analysed on an agarose gel and variants were distinguished by size: Variant 1 87bp, variant 2 98bp C. PCR with splice variant specific primers shows that all of the tested cell lines express the exon 1a as well as the exon 1b transcript variants. In order to detect both splice variants (left half of Figure) or only variant 2 (right half of Figure), PCRs were performed with the following primer combinations, ABP-F2/ABP-R2 (variant 1 and 2) or ABP-F2b/ABP-R2 (variant 2 specific) respectively. +, PCR with cDNA; -, PCR with total RNA.