Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
권호 표지
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Characterization and Evaluation of Melanocortin 4 Receptor (MC4R) Gene Effect on Pork Quality Traits in Pigs

돼지 Melanocortin 4 Receptor (MC4R) 유전자의 육질연관성 분석

  • Roh, Jung-Gun (College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Kim, Sang-Wook (College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Choi, Jung-Suk (College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Choi, Yang-Il (College of Agriculture, Life and Environment Sciences, Chungbuk National University) ;
  • Kim, Jong-Joo (School of Biotechnology, Yeungnam University) ;
  • Choi, Bong-Hwan (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science) ;
  • Kim, Tae-Hun (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science) ;
  • Kim, Kwan-Suk (College of Agriculture, Life and Environment Sciences, Chungbuk National University)
  • 노정건 (충북대학교 농업생명환경대학 축산학과) ;
  • 김상욱 (충북대학교 농업생명환경대학 축산학과) ;
  • 최정석 (충북대학교 농업생명환경대학 축산학과) ;
  • 최양일 (충북대학교 농업생명환경대학 축산학과) ;
  • 김종주 (영남대학교 생명공학부) ;
  • 최봉환 (농촌진흥청 국립축산과학원) ;
  • 김태헌 (농촌진흥청 국립축산과학원) ;
  • 김관석 (충북대학교 농업생명환경대학 축산학과)
  • Received : 2011.02.09
  • Accepted : 2012.01.31
  • Published : 2012.02.29
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Abstract

This study aimed to investigate the single nucleotide polymorphisms (SNPs) of the porcine MC4R gene and validate the effect of the MC4R genotype for marker assisted selection (MAS). Six amplicons were produced to analyze the entire base sequences of the porcine MC4R gene and six SNPs were detected (c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, and c.*430A>T). Linkage disequilibrium (LD) of the six SNPs was analyzed by performing haploid analysis. There was a perfect linkage disequilibrium in c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, and c.*430A>T. Only the c.892A>G (Asp298Asn) SNP showed a very low LD with an $r^2$ value of 0.028 and the D' value of 0.348. As a result, the two SNPs-c.707A>G (Arg236His) and c.892A>G (Asp298Asn)-were selected to extract the genotype frequencies from the 5 pig breeds by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotype analysis method. The SNP frequency of c.707A>G (Arg236His) indicated the presence of the A (His) allele only in Yorkshire, while the G allele was fixed in the KNP, Landrace, Berkshire, and Duroc. Association analysis was carried out in 484 pigs with the c.707A>G (Arg236His) SNP and the meat quality traits of four different pig cross populations: a significant association was noted in crude fat, sirloin moisture, meat color, and the degree of red and yellow coloration. The frequency of the c.892A>G(Asp298Asn) SNP genotype varied among the breeds; while Duroc showed the highest frequency of the A (Asn) allele, KNP showed the highest frequency of the G (Asp) allele. Association analysis was carried out in 1126 pigs with the c.892A>G (Asp298Asn) SNP and the meat quality traits of four pig populations: a highly significant linkage was noted in the back-fat thickness (P<0.002). It was found that the back-fat thickness was higher in individuals with the AA genotype than in those with the AG or GG genotype. Thus, in this study, we verified that the c.892A>G (Asp298Asn) SNP in the pig MC4R gene has a sufficient effect as a gene marker for MAS in Korean pork industry.

본 연구는 한국재래돼지의 MC4R 유전자 내의 단일염기변이들을 규명하고 그 유전자형 효과가 유전자표지인자를 이용한 선발(Marker assisted selection, MAS)에 활용 가능한지를 검증하기 위해서 수행되었다. 한국재래돼지의 MC4R 유전자 총 염기서열을 분석하기 위해 6개의 Primer들을 이용하여 증폭산물을 생성하였으며, 염기서열분석을 통해 총 6개(c.-780C>G, c-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, c.*430A>T)의 단일염기변이를 발견하였다. 한국재래돼지 MC4R 유전자내의 총 6개의 단일염기변이들간의 연관불균형과 반수체 분석을 통해 단일염기변이들간의 연관성을 분석하였으며, c.-780C>G, c-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His와 c.*430A>T는 완전한 연관불균형을 이루고 있었고, c.892A>G(Asp298Asn) 단일염기변이만 $r^2$-value가 0.028, D'-value가 0.348로 연관불균형 정도가 매우 낮았다. c.707A>G (Arg236His)와 c.892A>G (Asp298Asn) 단일염기변이들을 선발하여 PCR-RFLP 유전자형 분석방법을 이용해 돼지 5품종간의 유전자형 빈도를 추정한 결과, c.707A>G (Arg236His) 단일염기변이는 요크셔 품종 집단에서 오직 A (His) 대립유전자를 관찰할 수 있었으며, 나머지 한국재래돼지, 랜드레이스, 버크셔와 듀록 품종에서는 G 대립유전자의 고정으로 나타났다. c.707A>G 단일염기변이와 육질형질을 484두에서 연관성 분석을 실시한 결과, 조지방, 등심 내의 수분, 육색, 적색도 그리고 황색도 등에서 유의적인 연관성을 관찰할 수 있었다. c.892A>G (Asp298Asn) 단일염기변이의 유전자형 빈도는 품종별로 차이가 났으며, A (Asn) 대립유전자의 빈도가 가장 높은 품종은 듀록으로 나타났고, G (Asp) 대립유전자의 빈도가 가장 높은 품종은 한국재래돼지로 조사되었다. c.892A>G (Asp 298Asn) 단일염기변이와 돼지 4 집단의 육질형질을 1,126두에서 분석한 결과, 등지방두께에 고도의 유의적인 효과를 관찰할 수 있었다(P<0.002). AA 유전자형을 가진 개체가 AG나 GG 유전자형을 가진 개체보다 등지방두께가 두꺼운 것을 확인할 수 있었다. 본 연구의 결과를 통해 MC4R 유전자 내의 c.892A>G (Asp298Asn) 단일염기변이는 돼지의 선발개량에 유전자표지인자로서 충분한 효과가 있음을 검증하였다.

Keywords

References

  1. Barrett, J. E., Fry, B., Maller, J. and Daly, M. J. 2005. HAPLOVIEW: analysis and visualization of LD and haplotype maps. Bioinformatics. 21:263-265. https://doi.org/10.1093/bioinformatics/bth457
  2. Bruun, C. S., Jørgensen, C. B., Nielsen, V. H., Andersson. L. and Fredholm, M. 2006. Evaluation of the porcine melanocortin 4 receptor (MC4R) gene as a positional candidate for a fatness QTL in a cross between Landrace and Hampshire. Anim. Genet. 37:359-362. https://doi.org/10.1111/j.1365-2052.2006.01488.x
  3. Ciobanu, D., Bastiaansen, J., Malek, M., Helm, J., Woollard, J., Plastow, G. and Rothschild, M. F. 2001. Evidence for new alleles in the protein kinase adenosine monophosphate-activated gamma ${\gamma}3$-subunit gene associated with low glycogen content in pig skeletal muscle and improved meat quality. Genetics. 159:1151-1162.
  4. Ciobanu, D., Bastiaansen, J., Lonergan, S. M., Thomsen, H., Plastow, G. and Rothschild, M. F. 2004. New alleles in calpastatin gene are associated with meat quality traits in pigs. J. Anim. Sci. 82:2829-2839. https://doi.org/10.2527/2004.82102829x
  5. Choi, Y. S., Park, B. Y., Lee, J. M. and Lee, S. K. 2005. Comparison of carcass and meat quality characteristics between Korean native black pigs and commercial crossbreed pigs. Korean J. Food Sci. 25:322-327.
  6. Cho, S. H., Park, B. Y., Kim, J. H., Kim, M. J., Seong, P. N., Kim, Y. J., Kim, D. H. and Ahn, C. N. 2007. Carcass yields and meat quality by live weight of Korean native black pigs. Korea. J. Anim. Sci. Tech. 49:523-530. https://doi.org/10.5187/JAST.2007.49.4.523
  7. Fujii, J. K., Otsu, F., Zorzato, S., de Leon, V. K., Khanna, J. E., Weiler, P. J., O'Brien, D. H. and MacLennan, D. H. 1991. Identification of a mutation in porcine ryanodine receptor associated with malignant hyperthermia. Science. 253:448-451. https://doi.org/10.1126/science.1862346
  8. Farooqi, I. S., Keogh. J. M., Yeo, G. S., Lank, E. J., Cheetham, T. and O'Rahilly, S. 2003. Clinical spectrum of obesity and mutations in the melanocrtin-4 receptor gene. N. Engl. J. Me. 348:1085-1089. https://doi.org/10.1056/NEJMoa022050
  9. Fan, B., Onteru, S. K., Plastow, G. S. and Rothschild, M. F. 2009. Detailed characterization of the porcine MC4R gene in relation to fatness and growth. Anim. Genet. 40:401-409. https://doi.org/10.1111/j.1365-2052.2009.01853.x
  10. Han, S. H., Lee, S. S., Ko, M. S., Seong, P. N., Park, B. Y. and Cho, I. C. 2007. Effects of a porcine MC4R polymorphism (892G>A) on carcass traits in commercial pigs. Korea. J. Anim. Sci. Tech. 49:569-576. https://doi.org/10.5187/JAST.2007.49.5.569
  11. Houston, R. D., Cameron, N. D. and Rance, K. A. 2004. A melanocortin-4 receptor (MC4R) polymorphism is associated with performance traits in divergently selected Large White pig populations. Anim. Genet. 35:386-390. https://doi.org/10.1111/j.1365-2052.2004.01182.x
  12. Jin, S. K., Kim, I. S., Hur, S. J., Hah, K. H. and Kim, B. W. 2005. Physico-chemical characteristics with market weight in Korean native and landrace crossbred pigs. Korean J. Intl. Agri. 17: 182-187.
  13. Jin, S. K., Kim, I. S., Hur, S. J., Kim, S. J. and Jeong, K. J. 2006. The influence of pig breeds on qualities of loin. Korea. J. Anim. Sci. Tech. 48:747-758. https://doi.org/10.5187/JAST.2006.48.5.747
  14. Jokubka, R., Maak, S., Kerziene, S. and Swalve, H. H. 2006. Association of a melanocortin 4 receptor (MC4R) polymorphism with performance traits in Lithuanian White pigs. J. Anim. Breed. Genet.123:17-22. https://doi.org/10.1111/j.1439-0388.2006.00559.x
  15. Kim, E. H., Choi, B. H., Kim, K. S., Lee, C. K., Cho, B. W., Kim, T. H. and Kim, J. J. 2007. Detection of Mendelian and parentof- origin quantitative trait loci in a cross between Korean native pig and Landrace I. growth and body composition traits. Asian-Aust. J. Anim. Sci. 19:1702-1705.
  16. Kim. K. S., Larsen, N. J., Short, T., Plastow, G. and Rothschild, M. F. 2000a. A missense variant of the porcine melanocortin-4 receptor (MC4R) gene is associated with fatness, growth, and feed intake traits. Mamm. Genome. 11:131135. https://doi.org/10.1007/s003350010025
  17. Kim, K. S., Larsen, N. J. and Rothschild, M. F. 2000b. Linkage and physical mapping of the porcine melanocortin-4 receptor (MC4R) gene. J. Anim. Sci. 78:791792. https://doi.org/10.2527/2000.783791x
  18. Kim, K. S., Shin, H. Y., Lee, J. J., Hong, S. K., Choi, B. W., Kim, T. H., Lee, H. K. and Cho, B. W. 2005. Investigation of Porcine Melanocortin-4 Receptor (MC4R) Polymorphism on Economic Traits. Korea. J. Life. Sci. 15:968-971. https://doi.org/10.5352/JLS.2005.15.6.968
  19. Kim, M. J., Oh, J. D., Cho, G. H., Lee, J. H., Lee, S. S., Hong, Y. S., Jeon, J. K., Jeon, G. J. and Lee, H. K. 2006. Association between economic traits and candidate gene polymorphism in Korean native pig and Duroc. Korean. J. Emd. Trans. 21:73- 280.
  20. Kim, S. S., Kim, J. R., Moon, J. K., Choi, B. H., Kim, T. H., Kim, K. S., Kim, J. J. and Lee, C. K. 2009. Transcriptional alteration of p53 related processes as a key factor for skeletal muscle characteristics in Sus scrofa. Mol. Cells. 28:565-573. https://doi.org/10.1007/s10059-009-0159-z
  21. Kim, S. W., Roh, J. G., Cho, Y. I., Choi, B. H., Kim, T. H., Kim, J. J. and Kim, K. S. 2010. Development of Optimal Breeding Pigs Using DNA Marker Information. Genomics Inform. 8:81-85. https://doi.org/10.5808/GI.2010.8.2.081
  22. Li, S., Li, X., Choi, B. W., Lee, C. K., Cho, B. W., Kim. J. J. and Kim, K. S. 2009. Linkage disequilibrium and gene expression analyses of IGF2 gene in Korean native pigs. Korea. J. Anim. Sci. Tech. 51:15-24. https://doi.org/10.5187/JAST.2009.51.1.015
  23. Li, X., Lee, C. K., Choi, B. W., Kim, T. H., Kim, J. J. and Kim, K. S. 2010. Quantitative gene expression analysis on chromosome 6 between Korean native pigs and Yorkshire breeds for fat deposition. Genes Genom. 32:385-393. https://doi.org/10.1007/s13258-010-0009-6
  24. Meidtner, K., Wermter, A. K., Hinney, A., Remschmidt, H., Hebebrand, J. and Fries, R. 2006. Association of the melanocortin 4 receptor with feed intake and daily gain in F2 Mangalitsa x Pietrain pigs. Anim. Genet. 37:245-247. https://doi.org/10.1111/j.1365-2052.2006.01414.x
  25. Moon, J. K., Kim, K. S., Kim, J. J., Choi, B. H., Cho, B. W., Kim, T. H. and Lee, C. K. 2009. Differentially expressed transcripts in adipose tissue between Korean native pig and Yorkshire breeds. Anim. Genet. 40:115-118. https://doi.org/10.1111/j.1365-2052.2008.01798.x
  26. Oh, H., Kim, H., Yang, H., Lee, J., Joo, Y. and Kim, C. 2008. Comparison of meat quality characteristics between crossbreeds. Korean J. Food Sci. Ani. Resour. 28:171-180. https://doi.org/10.5851/kosfa.2008.28.2.171
  27. Park, H. B., Carlborg, O., Marklund, S. and Andersson, L. 2002. Melanocortin-4 receptor (MC4R) genotypes have no major effect on fatness in a Large White x Wild Boar intercross. Anim. Genet. 33:155-157. https://doi.org/10.1046/j.1365-2052.2002.00824.x
  28. Rothschild, M. F., Jacobson, C. and Vaske, D. A. 1996. The estrogen receptor locus is associated with a major gene influencing litter size in pigs. Proc. Natl. Acad. Sci. 93:201205. https://doi.org/10.1073/pnas.93.1.201
  29. Stephens, M., Smith, N. and Donnelly, P. 2001. A new statistical method for haplotype reconstruction from population data. Am. J. Hum. Genet. 69:978989.
  30. Stachowiak, M., Szydlowski, M., Obarzanek-Fojt, M. and Switonski, M. 2006. An effect of a missense mutation in the porcine melanocortin-4 receptor (MC4R) gene on production traits in Polish pig breeds is doubtful. Anim. Genet. 37:55-57. https://doi.org/10.1111/j.1365-2052.2005.01373.x
  31. Van den Maagdenberg, K., Stinckens, A., Claeys, E., Seynaeve, M., Clinquart, A., Georges, M., Buys, N. and De, Smet. S. 2007. The p.Asp298Asn missense mutation in the porcine melanocortin- 4 receptor (MC4R) gene can be used to affect growth and carcass traits without an effect on meat quality. Animal. 1:10891098.