Molecular Cloning And Analyzing Of Yak CAPN1 And CAPN2 CDNA

Molecular cloning and analyzing of yak CAPNland CAPN2 cDNACAPN1and CAPN2 genes are novel genes cloned recently. They were investigated as potential candidate genes affecting meat tenderness. The products of these two genesu,-calpain and m-calpain plays important roles in the tenderization process of meat and the allelic variation of these genes has potential impacts on the tenderness of meat. In this study The total RNAs were extracted from muscle tissues of yak. RT-PCR was performed to synthesize cDNA. According to the CAPN1 gene cDNA sequences of bovine, porcine and human published in GeneBank, the nested primers were designed to screen cDNA by performing nested PCR. The partial cDNAs of CAPN1 and CAPN2 genes were cloned and were ligated to pGEM-T-easy vector. DH5a was transformed by pGEM-T-easy vector. The positive clones were selected and plasmids extracted from them were identified by restriction endonuclease and sequencing.the partial nucleotide sequences of yak CAPN1 cDNA with 224 bp in length was identified, meanwhile , the partial nucleotide sequences of yak CAPN2 cDNA with 378 bp in length was also identified unexpectedly. The homology of these two nucleotide sequences with their counterpart of CAPN1 and CAPN2 genes were compared. The identity of CAPN1 nucleotide sequences was 98% between yak and cow, 89% between yak and human, 89% between yak and macaca, 83% between yak and mouse, 96% between yak and sheep, 81% between yak and rat. The identity of CAPN2 nucleotide sequences 98% between yak and sheep, 92% between yak and rabbit, 91% between yak and human, 90% between yak and macaca, 88% between yak and mouse. The nucleotide sequences of CAPN1 and CAPN2 genes between different species were comparatively conservative. Molecular cloning and analyzing of yak CAPN1 and CAPN2 partial cDNAs has made substantial preparations for cloning the complete nucleotide sequences of yak CAPN1 and CAPN2 genes.