Figure 3.

Production and analysis of F₁ generation hNGF-TG pigs. (A) EGFP expression in F₁ hNGF-TG pigs produced by mating the no. 5 F₀ hNGF-TG Duroc boar with WT Yorkshire sows (with dominant white coat color). (B) Southern blot analysis of transgene integration patterns in the genome of F₁ hNGF-TG pigs following EcoR V enzyme digestion. M: markers. N: negative control. P: positive controls using pmPSP-hNGF-EGFP plasmid as sample DNA. WT: wild-type piglets. 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30 are the ID no. of 10 offspring delivered by a WT sow mated with the no. 5 F₀ hNGF-TG boar. #1, #2, #3, and #4 are the four copies of transgene carried by no. 5 F₀ hNGF-TG boar. (C,D) The transgene integration site identified by inverse PCR in F₁ hNGF-TG pig no. 26 carrying the #1 copy of transgene, and no. 24 and no. 21 carrying the #2 copy of transgene, respectively. (E) hNGF concentration in oral saliva collected at rest time of no. 22, no. 24, no. 27, and no. 30 F₁ hNGF-TG pigs at the age of 5 months. (F) Average hNGF and pNGF protein concentrations in different tissues of WT control pigs and F₁ hNGF-TG pigs no. 22, no. 24, and no. 27 sacrificed at the age of 5 months. Sm: submandibular gland. Sl: sublingual gland. Pa: parotid gland. He: heart. Fa: fat. Lu: lung. Mu: muscle. ** means the difference between two groups reached statistical significance at p < 0.01.