Figure 4

Molecular analysis of the progeny from 'remobilized' frog 12M2♀5. Southern blot analysis was performed on genomic DNA harvested from individual tadpoles and digested with either BamHI (a) or BglII (b) and probed with a radiolabeled GFP probe to demonstrate the inheritance of novel transposon integration events. EPTS LM-PCR was used to amplify the genomic sequences flanking the 5' end of the Tol2 transposon. The amplified bands were resolved on agarose gels (c) and subcloned prior to sequence analysis. Sequence analysis (d) revealed that the novel remobilization events were the result of transposition as the insertion sites were flanked by the expected 8 bp target site duplications (TSD, underlined). The flanking genomic sequence is depicted by the capitalized text and the Tol2 sequence is shown in lowercase italics. The fol integration event was mapped to the same scaffold as the parental (hbr) integration site and represents a 'local hopping' transposition.