Hostname: page-component-76fb5796d-22dnz Total loading time: 0 Render date: 2024-04-26T08:40:29.424Z Has data issue: false hasContentIssue false
  • English
  • Français

Comparison of three PCR-based methods to detect a Piedmontese cattle point mutation in the Myostatin gene

Published online by Cambridge University Press:  01 June 2009

A. Pozzi ,
G. Bongioni  and
A. Galli
A. Pozzi*
Affiliation:
Istituto Sperimentale Italiano Lazzaro Spallanzani, Loc. La Quercia – 26027 Rivolta d’Adda, Cremona, Italy
G. Bongioni
Affiliation:
Istituto Sperimentale Italiano Lazzaro Spallanzani, Loc. La Quercia – 26027 Rivolta d’Adda, Cremona, Italy
A. Galli
Affiliation:
Istituto Sperimentale Italiano Lazzaro Spallanzani, Loc. La Quercia – 26027 Rivolta d’Adda, Cremona, Italy
*
E-mail: anna.pozzi@istitutospallanzani.it
Get access

Abstract

Despite many recent advances in single nucleotide polymorphism (SNP) genotyping technologies, there is still the need for methodologies with a reasonable throughput. In this study, we compared three PCR-based methods for SNP detection: (1) a conventional PCR-based allele detection system with fluorescent genotyping technology, (2) a SNaPshot methodology by single nucleotide primer extension and, (3) a real-time PCR-based method by allele-specific minor groove-binder probes. These three methodologies were used to analyze 104 meat samples of a particular Italian cattle breed known for producing excellent quality meat and for a characteristic increased development of muscle mass, caused by a point mutation (C313Y) in the Myostatin gene. The analysis revealed 98 samples to be homozygous (mh/mh) and five to be heterozygous (mh/+) for the mutation whereas one sample resulted to be homozygous for the wild type (+/+). The results obtained with the three different assays were consistent. Overall, all three methodologies proved to be efficient for allelic discrimination studies; however, real-time PCR was faster and allowed to genotype up to 96 samples in a single step, minimizing the number of steps required for samples manipulation.

Keywords

cattleMyostatin geneSNP detectiongenotyping
Type
Full Paper
Information
animal , Volume 3 , Issue 6 , June 2009 , pp. 773 - 778
Copyright
Copyright © The Animal Consortium 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Antoniou, F, Grosz, M 1999. PCR based detection of bovine myostatin Q204X mutation. Animal Genetics 30, 225–244.CrossRefGoogle ScholarPubMed
Bellinge, RHS, Liberles, DA, Iaschi, SPA, O’Brien, PA, Tay, GK 2005. Myostatin and its implications on animal breeding: a review. Animal Genetics 36, 1–6.CrossRefGoogle ScholarPubMed
Capoferri, R, Bongioni, G, Galli, A, Aleandri, R 2006. Genetic control of conventional labeling through the bovine meat chain production by single nucleotide polymorphisms (SNPs) using real-time PCR. Journal of Food Protection 69, 1971–1977.CrossRefGoogle ScholarPubMed
Casas, E, Keele, JW, Shackelford, SD, Koohmaraie, M, Sonstegard, TS, Smith, TPL, Kappes, SM, Stone, RT 1998. Association of the muscle hypertrophy locus with carcass traits in beef cattle. Journal of Animal Science 76, 468–473.CrossRefGoogle ScholarPubMed
Dunner, S, Miranda, ME, Amigues, Y, Cañon, J, Georges, M, Hanset, R, Williams, J, Ménissier, F 2003. Haplotype diversity of the myostatin gene among beef cattle breeds. Genetic Selection Evolution 35, 103–118.CrossRefGoogle ScholarPubMed
Fahrenkrug, SC, Casas, E, Keele, JW, Smith, TPL 1999. Technical note: direct genotyping of the double-muscling locus (mh) in Piedmontese and Belgian Blue cattle by fluorescent PCR. Journal of Animal Science 77, 2028–2030.CrossRefGoogle ScholarPubMed
Grobet, L, Martin, LJ, Poncelet, D, Pirottin, D, Brouwers, B, Riquet, J, Schoeberlein, A, Dunner, S, Ménissier, F, Massabanda, J, Fries, R, Hanset, R, Georges, M 1997. A deletion in the bovine myostatin gene causes the double-muscled phenotype in cattle. Nature Genetics 17, 71–74.CrossRefGoogle ScholarPubMed
Kambadur, R, Sharma, M, Smith, TPL, Bass, JJ 1997. Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle. Genome Research 7, 910–915.CrossRefGoogle ScholarPubMed
Lee, SJ, McPherron, AC 2001. Regulation of myostatin activity and muscle growth. Proceedings of the National Academy of Sciences of United States of America 9, 9306–9311.CrossRefGoogle Scholar
McPherron, AC, Lee, SJ 1997. Double muscling in cattle due to mutations in the myostatin gene. Proceedings of the National Academy of Sciences of United States of America 94, 12457–12461.CrossRefGoogle ScholarPubMed