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      Evaluation of single-nucleotide polymorphisms in CAPN1 for association with meat tenderness in cattle.

      Journal of animal science
      Alleles, Animals, Base Sequence, Calpain, genetics, Cattle, physiology, Exons, Female, Genotype, Haplotypes, Introns, Male, Meat, standards, Polymerase Chain Reaction, veterinary, Polymorphism, Single Nucleotide, Quantitative Trait Loci, Sequence Alignment

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          Abstract

          Micromolar calcium activated neutral protease (CAPN1) was evaluated as a candidate gene for a quantitative trait locus (QTL) on BTA29 affecting meat tenderness by characterization of nucleotide sequence variation in the gene. Single-nucleotide polymorphisms (SNP) were identified by sequencing all 22 exons and 19 of the 21 introns in two sires (Piedmontese x Angus located at the U.S. Meat Animal Research Center in Clay Center, NE; Jersey x Limousin located at AgResearch in New Zealand) of independent resource populations previously shown to be segregating meat tenderness QTL on BTA29. The majority of the 38 SNP were found in introns or were synonymous substitutions in the coding regions, with two exceptions. Exons 14 and 9 contained SNP that were predicted to alter the protein sequence by the substitution of isoleucine for valine in Domain III of the protein, and alanine for glycine in Domain II of the protein. The resource populations were genotyped for these two SNP in addition to six intronic polymorphisms and two silent substitutions. Analysis of genotypes and shear force values in both populations revealed a difference between paternal CAPN1 alleles in which the allele encoding isoleucine at position 530 and glycine at position 316 associated with decreased meat tenderness (increased shear force values) relative to the allele encoding valine at position 530 and alanine at position 316 (P < 0.05). The association of maternal alleles with meat tenderness phenotypes is consistent with the hypothesis of CAPN1 as the gene underlying the QTL effect in two independent resource populations and presents the possibility of using these markers for selective breeding to reduce the numbers of animals with unfavorable meat tenderness traits.

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          Consed:A Graphical Tool for Sequence Finishing

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            The role of Ca(2+)-dependent proteases (calpains) in post mortem proteolysis and meat tenderness.

            This manuscript summarizes research results from our laboratory regarding the role of endogenous proteases in post mortem proteolysis resulting in meat tenderization. Proteolysis of key myofibrillar proteins is the principal reason for ultrastructural changes in skeletal muscle associated with meat tenderization. Proteases should have the following characteristics to be considered as possible candidates for bringing about post mortem changes: i) to be located within skeletal muscle cells; ii) to have access to the substrate ie, myofibrils); and iii) to be able to hydrolyze the same proteins that are degraded during post mortem storage. Of the proteases located within skeletal muscle cells and thus far characterized, only calpains have all of the above characteristics. Numerous experiments conducted in our laboratory have indicated that the calcium-dependent proteolytic system (calpains) is responsible for post mortem proteolysis. Some of this evidence includes: 1) incubation of muscle slices with buffer containing Ca2+ accelerates post mortem proteolysis; 2) incubation of muscle slices with Ca2+ chelators inhibits post mortem proteolysis; 3) infusion or injection of carcasses with a solution of calcium chloride accelerates post mortem proteolysis and the tenderization process such that post mortem storage beyond 24 h to ensure meat tenderness is no longer necessary; 4) infusion of carcasses with zinc chloride, a potent inhibitor of calpains, blocks post mortem proteolysis and the tenderization process; and 5) feeding a beta-adrenergic agonist to lambs results in a reduction of the proteolytic capacity of the calpain system, which leads to a decreased rate of post mortem proteolysis and produces tough meat.(ABSTRACT TRUNCATED AT 250 WORDS)
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              Bovine CAPN1 maps to a region of BTA29 containing a quantitative trait locus for meat tenderness.

              Micromolar calcium activated neural protease (CAPN1) was investigated as a potential candidate gene for a quantitative trait locus (QTL) on BTA29 affecting meat tenderness. A 2,948-bp bovine cDNA containing the entire coding region of the gene was obtained, showing 91% identity to human CAPN1. The 716 AA protein predicted from this sequence shows 97% similarity (95% identity) to the 714 AA human protein. Analysis of the gene structure revealed that CAPN1 mRNA is encoded by at least 19 exons, and 11,055 bp of the gene were sequenced, including 17 introns. Two single nucleotide polymorphisms (SNP) were detected in intron 12 and were used to map bovine CAPN1 to the telomeric end of the BTA29 linkage group. This approximately coincides with the position of the QTL, demonstrating that CAPN1 protease is a positional candidate gene potentially affecting variation in meat tenderness in a bovine resource mapping population.
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