Figure 1: Generation and characterization of PEPCK-Cmus transgenic pigs.
(a) Schematic diagram of the transgenic vector (pZT52). (b) Photograph of eleven cloned PEPCK-Cmus Tibetan mini pigs. (c) PCR analysis of genomic DNA from ear biopsies from the eleven cloned pigs. Six pigs (number T01, T03, T04, T05, T09 and T11) showed the expected 1,000 bp band. T01-T11, cloned pigs; M, marker; WT, wild type Tibetan mini pig as negative control; NTC, no template control. The full-length gel for this PCR analysis is presented in Supplementary Figure 1. (d) Real-time PCR analysis of PEPCK-C mRNA levels of skeletal muscle tissue from different anatomical locations (psoas, foreleg, hind leg, gluteus) of the transgenic and control pigs. PEPCK-C expression in wild type piglets’ liver tissue was selected as a positive control. Results were normalized to GAPDH. (e) Western blot analysis of PEPCK-C in skeletal muscle tissue from different anatomical locations (waist, foreleg, hind, gluteal) of the transgenic and control pigs. PEPCK-C expression in wild-type piglet liver tissue was selected as a positive control. α-Tubulin protein was used as an internal reference to demonstrate equal amounts of proteins were loaded. The full-length blots for this Western analysis are presented in Supplementary Figure 2 (PEPCK-C expression) and Supplementary Figure 3 (α-Tubulin expression).
