Cloning and SNPs analysis of MC1R gene of hybrid F2 generation of Landace × Bama minipig

ObjectiveThe influence of base mutation of melanocortin Ⅰ receptor (MC1R) gene on segregation of coat color was studied to provide references for its breeding and genetic mechanism of coat color. MethodPrimers were designed according to the MC1R gene coding sequence ( AF326520 ) obtained from GenBank . The total DNA in blood of 70 hybrid F2 generation of Landace ×Bama minipig was used as the template for PCR. The coding sequence of MC1R was amplified and cloned by PCR. SNPs analysis of MC1R gene was performed using PCR-SSCP. ResultMC1R gene coding sequence (CDS) of the hybrid F2 generation of Landace and Bama minipig was 963 bp, in which similarity was 99.37%, compared with its reference sequence. MC1R gene coding sequences (CDS) in the hybrid F2 generation of Landace and Bama minipig had six mutations compared to sequences of MC1R gene from GenBank. The base 284G→A caused amino acid 95Val→Met and 306T→C led to 102Leu→Pro , except 4 synonymous mutations , which were 6C→T, 51G→A, 364T→C and 730G→A . PCR-SSCP analysis showed that MC1R displayed no polymorphism. Con-clusionMC1R gene has polymorphism in the hybrid F2 generation of Landace×Bama minipig and its genotype is ED1,but none of polymorphism is found from the CDS of MC1R gene in the hybrid F2 generation of Landace×Bama minipig, in which coat color was different , so coat color polymorphism can not be simply explained by polymorphism of MC1R.