DOI QR코드

DOI QR Code

Identification of Korean Native Pork Using Breed-Specific DNA Marker of KIT Gene

  • Chung, Eui-Ryong (Division of Animal Science and Resources, College of Life Science and Natural Resources, Sangji University) ;
  • Chung, Ku-Young (Division of Animal Science and Resources, College of Life Science and Natural Resources, Sangji University)
  • Received : 2010.02.26
  • Accepted : 2010.05.14
  • Published : 2010.06.30

Abstract

Accurate methods for the identification of closely related species or breeds in raw and processed meats must be developed in order to protect both consumers and producers from mislabeling and fraud. This paper describes the development of DNA markers for the discrimination and improvement of Korean native pig (KNP) meat. The KIT gene is related to pig coat color and is often used as a candidate marker. A 538 bp fragment comprising intron 19 of the pig KIT gene was amplified by PCR using specific primers, after which the PCR amplicons of a number of meat samples from KNP and three major improved breeds (Landrace, Duroc and Yorkshire) were sequenced in order to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of two nucleotide substitutions, g.276G>A and g.295A>C, between KNP and the improved pig breeds. Digestion of KIT amplicons with AccII enzyme generated characteristic PCR-RFLP profiles that allowed discrimination between meats from KNP and improved pig. KNP showed three visible DNA bands of 264/249, 199, and 75 bp, whereas DNA bands of 249, 199, and 90 bp were detected in the three improved pig breeds. Therefore, the 75 bp DNA fragment was specific only to KNP, whereas the 90 bp DNA fragment was specific to the improved breeds. The breed-specific DNA markers reported here that target the KIT gene could be useful for the identification of KNP meat from improved pig meats, thus contributing to the prevention of falsified breed labeling.

Keywords

KIT gene;PCR-RFLP;breed-specific DNA marker;identification of KNP meat

References

  1. Calvo, J. H., Zaragoza, P., and Osta, P. (2001) Technical note: A quick and sensitive method to identify pork in processed and unprocessed food by PCR amplification of a new specific DNA fragment. J. Anim. Sci. 79, 2108-2212.
  2. 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. Korean J. Anim. Aci. Technol. 49, 523-530. https://doi.org/10.5187/JAST.2007.49.4.523
  3. Chung, E. R., Kim, W. T., Kim, Y. S., and Han, S. K. (2000) Identification of Hanwoo meat using PCR-RFLP marker of MC1R gene associated with bovine coat color. Korean J. Anim. Sci. Technol. 42. 379-380.
  4. Colgan, S., O'Brien, L., Maher, M., Shilton, N., McDonnell, K., and Ward, S., (2001) Development of a DNA-based assay for species identification in meat and bone meal. Fleischwirtschaft. 34, 409-414.
  5. Fajardo, V., Gonzalez, I., Lopez-Calleja, I., Martin, I., Rojas, M., Garcia, T., Hernandez, P. E., and Martin, R. (2007) PCR identification of meats from chamois (Rupicapra rupicapra), Pyrenean ibex (Capra pyrenaica), and mouflon (Ovis ammon) targeting specific sequences from the mitochondrial D-loop region. Meat Sci. 76, 644-652. https://doi.org/10.1016/j.meatsci.2007.02.002
  6. Girish, P. S., Anjaneyulu, A. S. R., Viswas, K. N., Shivakumar, M., Anand, M., Patel, M., and Sharma, B. (2005) Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci. 70, 107-112. https://doi.org/10.1016/j.meatsci.2004.12.004
  7. Jin, S. K., Kim, C. W., Song, Y. M., Jang, W. H., Kim, Y. B., Yeo, J. S., Kim, J. W., and Kang, K. H. (2001) Physicochemical characteristics of longissimus muscle between the Korean native pig and Landrace. Korean J. Food Sci. Anim. Resour. 21, 142-148.
  8. Johansson Moller, M., Chaudhary, R., Hellmen, E., Hoyheim, B., and Chowdhary, B., (1996) Pigs with the dominant white coat color phenotype carry a duplication of the KIT gene encoding the mast/stem cell growth factor receptor. Mamm. Genome. 7, 822-830. https://doi.org/10.1007/s003359900244
  9. Kesmen, Z., Sahin, F., and Yetim, H. (2007) PCR assay for the identification of animal species in cooked sausages. Meat Sci. 77, 1-5.
  10. Kijas, J. M., Wales, R., Tornsten, A., Chardon, P., and Moller, M., (1998) Melanocortin receptor 1 (MC1R) mutations and coat color in pigs. Genetics. 150, 1177-1185.
  11. Klungland, H., Vage, D. I., Gomez-Raya, L., Adalsteinsson, S., and Lien, S., (1995) The role of melanocyte-stimulating hormone (MSH) receptor in bovine coat colour determination. Mamm. Genome. 6, 636-639. https://doi.org/10.1007/BF00352371
  12. Koh, M. C., Lim, C. H., Chua, S. B., Chew, S. T., and Phang, S. T. W. (1998) Random amplified polymorphic DNA (RAPD) fingerprints for identification of red meat animal species. Meat Sci. 48, 275-285. https://doi.org/10.1016/S0309-1740(97)00104-6
  13. Lee, S. K., Ju, M. K., Kim, Y. S., Kang, S. M., and Choi, Y. S. (2005) Quality comparison between Korean native black groun pork and modern genotype pork during refrigerated storage. Kor J. Food Sci. Ani. Resour. 25, 71-77.
  14. Matsunaga, T., Chikuni, K., Tanabe, R., Muroya, S., Nakai, H., Shibata, K., Yamada, J., and Shinmura, Y. (1998) Determination of mitochondrial cytochrome b gene sequence for red deer (Cervus elaphus) and the differentiation of closely related deer meats. Meat Sci. 49, 379-385. https://doi.org/10.1016/S0309-1740(97)00145-9
  15. Matsunaga, T., Chikuni, K., Tanabe, R., Muroya, S., Shibata, K., Yamada, J., and Shinmura, Y. (1999) A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Sci. 51, 143-148. https://doi.org/10.1016/S0309-1740(98)00112-0
  16. Marklund, S., Moller, M., Sandberg, K., and Andersson, L. (1999) Close association between sequence polymorphism in the KIT gene and the roan coat color in horses. Mamm. Genome. 10, 283-288. https://doi.org/10.1007/s003359900987
  17. Marklund, S., Kijas, R., Rodriguez-Martinez, H., Ronstrand, L., and Funa, K. (1998) Molecular basis for the dominant white phenotype in the domestic pig. Genome Res. 8, 826- 833.
  18. Meyer, R., Hoefelein, C., Luethy, J., and Candrian, U. (1995) Polymerase chain reaction-restriction fragment length polymorphism analysis: a simple method for species identification in food. J. AOAC Intl. 78, 1542-1551.
  19. Montiel-Sosa, J. F., Ruiz-Pesini, E., Montoya, J., Rocales, P., Lopez-Perez, M. J., and Perez-Martos, A. (2000) Direct and highly species-specific detection of pork meat and fat in meat products by PCR amplification of mitochondrial DNA. J. Agri. Food Chem. 48, 2829-2832. https://doi.org/10.1021/jf9907438
  20. Okumura, N., Kobayashi, E., Suzuki, H., Morozumi, T., Hamashima, N., and Mitsuhashi, T. (2000) Breed specific mutations in MC1R and KIT genes in pigs. Anim. Sci. J. 8, 222-234.
  21. Saez, R., Sanz, Y., and Toldra, F. (2004) PCR-based fingerprinting techniques for rapid detection of animal species in meat products. Meat Sci. 66, 659-665. https://doi.org/10.1016/S0309-1740(03)00186-4
  22. Sasazaki, S., Mutoh, H., Tsurifune, K., and Mannen, H. (2007) Development of DNA markers for discrimination between domestic and imported beef. Meat Sci. 77, 161-166. https://doi.org/10.1016/j.meatsci.2007.02.024
  23. Shin, S. C., Chung, K. Y., and Chung, E. R. (2006) Identification of meat species using PCR-RFLP marker of cytochrome b gene. Korean J. Food. Sci. Ani. Resour. 26, 375- 379.
  24. Giuffra, E., Evans G., Tornsten, A., Wales, R., Day A., Looft, H., Plastow, G., and Andersson, L. (1999) The belt mutation in pigs is an allele at the dominant white (I/KIT) locus. Mamm. Genome. 10, 1132-1136. https://doi.org/10.1007/s003359901178
  25. Martin, I., Garcia, T., Fajardo, V., Lopez-Calleja, I., Rojas, M., Hernandez, P. E., Gonzalez, I., and Martin, R. (2007) Mitochondrial markers for the detection of four duck species and the specific identification of Muscovy duck in meat mixtures using the polymerase chain reaction. Meat Sci. 76, 721- 729. https://doi.org/10.1016/j.meatsci.2007.02.013
  26. 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 crossbred pigs. Korean J. Food Sci. Ani. Resour. 25, 322-327.
  27. Kang, S. M., Kang, C. G., and Lee, S. K. (2007) Comparison of quality characteristics of korean native black pork and modern genotype pork during refrigerated storage after thawing. Korean J. Food. Sci. Ani. Resour. 27, 1-7. https://doi.org/10.5851/kosfa.2007.27.1.1
  28. Park, J. K., Shin, K. H., Shin, S. C., Chung, K. Y., and Chung, E. R. (2007) Identification of meat species using species- specific PCR-RFLP fingerprint of mitochondrial 12S rRNA gene. Korean J. Food Sci. Ani. Resour. 27, 209-215. https://doi.org/10.5851/kosfa.2007.27.2.209

Cited by

  1. Authentication of “mono-breed” pork products: Identification of a coat colour gene marker in Cinta Senese pigs useful to this purpose vol.184, 2016, https://doi.org/10.1016/j.livsci.2015.12.007