Fig. 3.

Southern blot hybridisation analysis to confirm transgenic animals and estimate of copies number. DNA (5.6  μg) isolated from transgenic animals TG252, TG1183, TG1036 and TG1040, respectively, carrying the pGAL-GFPBsd gene construct was digested with EcoRI and hybridised with a probe of 464 bp. The probe was a 32P-dATP-labelled PCR product of 464 bp formed on plasmid DNA (pGAL-GFPBsd) with the use of the first pair of primers used for screening of the transgene. The probe detects a 7.3-kb band that represents the pGAL-GFPBsd genetic construct (black arrow). The red arrow probably indicates the band of non-specific hybridisation of the probe. Lanes 1–4 contain DNA samples from transgenic animals (TG252, TG1183, TG1036, TG1040); lane 5 wild-type pig DNA as a negative control; lanes 6–10 titration of the transgene mixed with digested wild-type genomic DNA representing 133.12, 66.56, 33.28, 16.64 and 8.32 copies, respectively