Figure 3.

Analysis of MC1R gene of cloned Durocs with light black coat color. (A) Representative image of a cloned Duroc (#8) showing the light black coat color. (B) Quantification of MC1R mRNA level in ear skin among cloned Durocs with three different coat colors (red, dark black, and light black). (C) Sanger sequencing results of genome DNA showed that #27 had the heterozygous E^D1/e alleles, and #8 had the heterozygous E^D1/E⁺ alleles (one allele of MC1R was changed to E^D1 genotype and the other allele had a V164A substitution forming an E⁺ genotype). Sanger sequencing of cDNA showed an equal expression level for both E alleles in light black pigs, but a great higher expression for the E^D1 allele in dark black pigs. Red arrows point to SNP sites varied in E^D1 and e alleles. AA, amino acid position.