Fig. 1

Establishing transgenic donor pig fetal fibroblasts carrying the CreER^T2 inducible system for SCNT. (A) Schematic diagram of the TB-pTYR::Cre system for porcine melanoma model production. The cassette regulates melanocyte-specific expression of CreER^T2, further inducing recombination of the construct and expression of mutant pig TP53 and human BRAF. Arrows labeled with lower case letters represent primers for PCR analysis to confirm genome integration of the system (a: Oncogene cassette; b: CreER^T2) (B) FACS analysis of the GFP-positive population of transgenic donor fibroblasts carrying the CreER^T2 inducible system. (C) Representative image of GFP or DsRed expression in transgenic donor fibroblasts. Scale bar, 200 μm. (D) PCR analysis confirming genomic integration of the CreER^T2 inducible system in transgenic donor fibroblasts using genomic DNA derived from control and transgenic fibroblasts. Non-transgenic (TG), genomic DNA extracted from normal porcine cell; TG Donor, genomic DNA from the transgenic donor cells; Plasmid, TB-pTYR::Cre plasmid vector.