Molecular Cloning and Sequence Analysis of CAPN1 Gene in Yak

CAPN1 was investigated as a potential candidategene for a quantitative trait locus(QTL) affecting meat tenderness.According to the CAPN1 gene sequence of Cattle in GenBank,the specific primers were designed.Using the Tianzhu White yak cDNA as template,PCR amplifying,cloning and sequencing for every fragment.Using the BioEdit to splice sequences and a 2267bp yak cDNA of CAPN1 gene was obtained.Including a complete open reading frame and partical sequence of 3' end and 5' end UTR.Analysis of the gene revealed that the yak CAPN1 gene is encoded by 2151bp and the 716 AA potein predicted from this sequence.The nucleotide sequence and amino acid sequence of yak calpain were compared with those of cattle,pig,human,and mouse calpain.Identities for nucleotide and amino acid sequences were 99.3% and 99.4% for cattle,93.9% and 96.1% for pig,90.0% and 94.6% for human,and 85.5% and 89.0% for mouse calpain.Search using NCBI BLAST of yak 4 domains respectively found the 4 domains in the above 4 species have shown good conservative,the most conservative in domain IV( 96%).There are 14 nucleotide mutations between yak and cattle and 3 of that were predicted to alter the protein sequence,both of which occurred in the domain III.Construction of molecular phylogenetic tree shows:The cluster results coincided with the traditional taxonomy.