Original Scientific Article
Allele and genotype frequencies of the κappa-casein (CSN3) locus in Macedonian Holstein-Friesian cattle
Nikola Adamov * ,
Branko Atanasov ,
Ksenija Ilievska ,
Martin Nikolovski ,
Monika Dovenska ,
Vladimir Petkov ,
Toni Dovenski

Mac Vet Rev 2020; 43 (1): 45 - 54

10.2478/macvetrev-2020-0013

Received: 15 October 2019

Received in revised form: 13 January 2020

Accepted: 26 January 2020

Available Online First: 06 February 2020

Published on: 15 March 2020

Correspondence: Nikola Adamov, adamovn@fvm.ukim.edu.mk
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Abstract

The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P<0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.

Keywords: CSN3, genetic polymorphism, Holstein-Friesian cattle, kappa-casein, milk protein

References

  1. Azevedo, A.L.S., Nascimento, C.S., Steinberg, R.S., Carvalho, M.R.S., Peixoto, M.G.C.D., Teodoro, R.L. et al. (2008). Genetic polymorphism of the kappa-casein gene in Brazilian cattle. Gen. Mol. Res. 7(3): 623-630. https://doi.org/10.4238/vol7-3gmr428 PMid:18752189
  2. Pacheco Contreras, V.I., Lourenco Jaramillo, D.L., Parra Bracamonte, G.M., Martínez González, J.C., Sifuentes Rincón, A.M. (2011). Convenient genotyping of nine bovine κ-casein variants. Electron J Biotechnol. 14(4): 12-12. https://doi.org/10.2225/vol14-issue4-fulltext-10 
  3. Farrell Jr., H.M., Jimenez-Flores, R., Bleck, G.T., Brown, E.M., Butler, J.E., Creamer, L.K. et al. (2004). Nomenclature of the proteins of cow's milk-sixth revision. J Dairy Sci. 87, 1641-1674. https://doi.org/10.3168/jds.S0022-0302(04)73319-6 
  4. Mercier, J.C., Chobert, J.M., Addeo, F. (1976). Comparative study of the amino acid sequences of caseinomacropeptides from seven species. FEBS Letters 72(2): 208-214. https://doi.org/10.1016/0014-5793(76)80972-6
  5. Sandra, S., Alexander, M., Dalgleish, D.G. (2007). The rennet coagulation mechanism of skim milk as observed by transmission diffusing wave spectroscopy. J Colloid Interface Sci. 308, 364-373. https://doi.org/10.1016/j.jcis.2007.01.021 PMid:17266978      
  6. Gallagher, D.S., Schelling, C.P., Groenen, M.M.A., Womack, J.E. (1994). Confirmation that the casein gene cluster resides on cattle chromosome 6. Mammalian Genome 5(8): 524-524. https://doi.org/10.1007/BF00369327 PMid:7949742 
  7. Threadgill, D.W., Womack, J.E. (1990). Genomic analysis of the major bovine casein genes. Nucleic Acids Res. 18, 6935-6942. https://doi.org/10.1093/nar/18.23.6935 PMid:1979856 PMCid:PMC332753  
  8. Popescu, C. P., Long, S., Riggs, P., Womack, J., Schmutz, S., Fries, R., Gallagher, D. S. (1996). Standardization of cattle cariotype nomenclature: Report of the committee for the standardization of the cattle cariotype. Cytogenet Cell Genet. 74, 259-261. https://doi.org/10.1159/000134429 PMid:8976379              
  9. Hayes, H., Petit, E., Bouniol, C., Popescu, P. (1993). Localization of the α-S2-casein gene (CASAS2) to the homoeologous cattle, sheep, and goat chromosomes 4 by in situ hybridization. Cytogenetic and Genome Research 64(3-4): 281-285. https://doi.org/10.1159/000133593 PMid:8404055  
  10. Alexander, J.L., Stewart, F.A., Mackinlay, G.A., Kapelinskaya, V.T., Tkach, M.T., Gorodetsky, I.S. (1988). Isolation and characterization of the bovine κ-casein gene. Eur J Biochem. 178, 395-401. https://doi.org/10.1111/j.1432-1033.1988.tb14463.x PMid:3208764     
  11. Martin, P., Szymanowska, M., Zwierzchowski, L., Leroux, C. (2002). The impact of genetic polymorphisms on the protein composition of ruminant milks. Reproduction Nutrition Development 42(5): 433-459. https://doi.org/10.1051/rnd:2002036 PMid:12537255
  12. Caroli, A.M., Chessa, S., Erhardt, G.J. (2009). Milk protein polymorphisms in cattle: Effects on animal breeding and human nutrition. J Dairy Sci. 92, 5335-5352. https://doi.org/10.3168/jds.2009-2461 PMid:19841193 
  13. Damiani, G., Ferretti, L., Rognoni, G., Sgaramella, V. (1990). Restriction fragment length polymorphism analysis of the kappa casein locus in cattle. Anim Genet. 21, 107-114. https://doi.org/10.1111/j.1365-2052.1990.tb03214.x PMid:1974749            
  14. Prinzenberg, E.M., Krause, I., Erhardt, G. (1999). SSCP analysis at the bovine CSN3 locus discriminates six alleles corresponding to known protein variants (A, B, C, E, F, G) and three new DNA polymorphisms (H, I, A1). Anim Biotechnol. 10(1-2): 49-62. https://doi.org/10.1080/10495399909525921 PMid:10654430              
  15. Prinzenberg, E.M., Jianlin, H., Erhardt, G., (2008). Genetic variation in the κ-casein gene (CSN3) of Chinese yak (Bos grunniens) and phylogenetic analysis of CSN3 sequences in the genus Bos. J Dairy Sci. 91(3): 1198-1203. https://doi.org/10.3168/jds.2007-0746 PMid:18292276          
  16. Thompson, M.P., Farrell, Jr. H.M. (1974). Genetic variants of the milk proteins. In B. L. Larson and V. R. Smith (Eds.), Lactation, vol. III (pp. 109-134). Academic Press, New York, NY. https://doi.org/10.1016/B978-0-12-436703-6.50008-9
  17. Bech, A.M., Kristiansen, K.R. (1990). Milk protein polymorphism in Danish dairy cattle and the influence of genetic variants on milk yield. J Dairy Res. 57, 53-62. https://doi.org/10.1017/S0022029900026601 PMid:2312876   
  18. Ng-Kwai-Hang, K.F., Grosclaude, F. (2003). Genetic polymorphism of milk proteins. In: P.F. Fox, P.L.H. McSweeney (Eds.), Advanced Dairy Chemistry - 1 Proteins (pp. 739-816). Springer, Boston, MA. https://doi.org/10.1007/978-1-4419-8602-3_22
  19. Patel, R.K., Chauhan, J.B., Singh, M.K., Soni, K.J. (2007). Genotype and allele frequencies of κ-casein and β-lactoglobulin in Indian river buffalo bulls (Bubalus bubalis). Buffalo Bulletin 26, 63-66.
  20. Buchberger, J., Dovc, P. (2000). Lactoprotein genetic variants in cattle and cheese making ability. Food Technol Biotechnol. 38(2): 91-98.
  21. Bovenhuis, H., Van Arendonk, J.A.M., Korver, S. (1992). Associations between milk protein polymorphisms and milk production traits. J Dairy Sci. 75, 2549 - 2559. https://doi.org/10.3168/jds.S0022-0302(92)78017-5
  22. Pierce, B.A. (2012). Population Genetics. In: S. Winslow (Ed.), Genetics - a Conceptual Approach (pp. 669-696). W.H. Freeman and Company.      
  23. Hamilton M.B. (2009). Genotype Frequencies. In: Population Genetics (pp. 9-52). Wiley-Blackwell.              
  24. Snustad, D.P., Simmons, M.J. (2012). Population Genetics. In: K. Witt and L. Morris (Eds.), Principles of Genetics (pp. 634-655). John Wiley and Sons, Inc.    
  25. Jensen, H.B., Holland, J.W., Poulsen, N.A., Larsen, L.B. (2012). Milk protein genetic variants and isoforms identified in bovine milk representing extremes in coagulation properties. J Dairy Sci. 95, 2891-2903. https://doi.org/10.3168/jds.2012-5346 PMid:22612926            
  26. Van Eenennaam, A., Medrano, J.F. (1991). Milk protein polymorphism in California dairy cattle. J Dairy Sci. 74, 1730-1742. https://doi.org/10.3168/jds.S0022-0302(91)78336-7
  27. Alipanah, M., Kalashnikova, L.A. (2007). Influence of K-casein genetic variant on cheese making ability. J Anim Vet Adv. 6(7): 855-857.
  28. Aleandri, R., Buttazzoni, L.G., Schneider, J.C., Caroli, A., Davoli, R. (1990). The effects of milk protein polymorphisms on milk components and cheese-producing ability. J Dairy Sci. 73, 241-255. https://doi.org/10.3168/jds.S0022-0302(90)78667-5
  29. Pagnacco, G., Caroli, A. (1987). Effect of casein and β-lactoglobulin genotypes on renneting properties of milk. J Dairy Res. 54(4): 479-485. https://doi.org/10.1017/S0022029900025681
  30. Marziali, A.S., Ng-Kwai-Hang, K.F. (1986). Relationships between milk protein polymorphisms and cheese yielding capacity. J Dairy Sci. 69(5): 1193-1201. https://doi.org/10.3168/jds.S0022-0302(86)80523-9
  31. Kubarsepp, I., Henno, M., Viinalass, H., Sabre, D. (2005). Effects of k-casein and β-lactoglobulin genotypes on the milk rennet coagulation properties. Agricultural Research 3, 55-64. https://doi.org/10.1080/09064700500453377
  32. Ikonen, T., Ahlfors, K., Kempe, R., Ojala, M., Ruottinen, O. (1999). Genetic parameters for the milk coagulation properties and prevalence of noncoagulating milk in Finnish dairy cows. J Dairy Sci. 8, 205-214. https://doi.org/10.3168/jds.S0022-0302(99)75225-2  
  33. Di Stasio, L., Mariani, P. (2000). The role of protein polymorphism in the genetic improvement of milk production. Zoot Nutr Anim 26, 69-90.        
  34. Abdel Dayem, A.M.H., Mahmoud, Gh.M. K., Nawito, M.F., Ayoub, M.M., Darwish, F.S. (2009). Genotyping of kappa casein gene in Egyptian buffalo bulls. Livest Sci. 122, 286-289. https://doi.org/10.1016/j.livsci.2008.09.010
  35. Medrano, J.F., Aguilar-Cordova, E. (1990). Genotyping of bovine kappa-casein loci following DNA sequence amplification. Biotechnology 8, 144-146. https://doi.org/10.1038/nbt0290-144 PMid:1369440              
  36. Galila, A.E., Samah, F.D. (2008). A PCR-RFLP assay to detect genetic variants of kappa-casein gene in cattle and buffalo. Arab J Biotech. 11(1): 11-18.
  37. Ilie, D., Salajeanu, A., Magdin, A., Stanca, C., Vintila, C., Vintila, I., Gocza, E. (2007). Genetic polymorphism at the k-casein locus in a dairy herd of Romanian spotted and Brown of maramures breeds. Scientific Papers Animal Science and Biotechnologies 40(1): 101-106.
  38. Anggraeni, A., Sumantri, C., Farajallah, A., Andreas, E. (2010). Kappa-casein genotypic frequencies in Holstein-Friesian dairy cattle in West Java province. Media Peternakan 33(2): 61-67. https://doi.org/10.5398/medpet.2010.33.2.61
  39. Alipanah, M., Kalashnikova, L., Rodinov, G. (2005). Kappa-casein genotypic frequencies in Russian breeds Black and Red Pied cattle. Iran. J Biotechnol. 3(3): 191-194.     
  40. Sitkowska, B., Neja, W., Wisniewska, E. (2009). Relations between kappa-casein polymorphism (CSN3) and milk performance traits in heifer cows. J Cent Eur Agric. 9(4): 641-644.      
  41. Lukac, D., Vidovic, V., Nemes, Z. Stupar, M., Popovic-Vranjes, A. (2013). Genotypic frequencies of the β-lactoglobulin, κ-casein and transferrin in Serbian Holstein-Friesian dairy cattle. Mljekarstvo 63 (4): 203-210.
  42. Ivankovic, A., Ramljak, J., Dokso, A., Kelava, N., Konjacic, M., Paprika, S. (2011). Genetic polymorphism of β-lactoglobulin and k-casein of cattle breeds in Croatia. Mljekarstvo 61(4): 301-308. [in Croatian]              
  43. Soria, L.A., Iglesias, G.M., Huguet, M.J., Mirande, S.L. (2003). A PCR-RFLP test to detect allelic variants of the bovine kappa-casein gene. Anim Biotech. 14(1): 1-5. https://doi.org/10.1081/ABIO-120020180  PMid:12887174 
  44. Tanaskovska, R.B., Srbinovska, S., Andonov, S., Trojacanec, S., Nestoriovski, T., Popovski, T.Z. (2016). Genotipization of k-casein in Holstein-Friesian cattle in Macedonia and its association with some milk properties. Int J Agr Innov Res. 5(2): 266-270.           
  45. Ren, D.X., Miao, S.Y., Chen, Y.L., Zou, C.X., Liang, X.W., Liu, J.X. (2011). Genotyping of the k-casein and β-lactoglobulin genes in Chinese Holstein, Jersey and water buffalo by PCR-RFLP. J Genet. 90(1): 1-5. https://doi.org/10.1007/s12041-011-0048-z
  46. Ju, Z., Huang, J., Li, Q., Wang, H., Zhong, J., Wang, C. (2011). The polymorphisms of κ-casein gene and their associations with milk production traits and expression analysis in Chinese Holstein cattle. Afric Jour Biotech. 10(62): 13368-13375. https://doi.org/10.5897/AJB10.1886
  47. Doosti, A., Arshi, A., Vatankhah, M., Amjadi, P. (2011). Kappa-casein gene polymorphism in Holstein and Iranian native cattle by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Afric Jour Biotech. 10(25): 4957-4960.         
  48. Toorchi, M., Tahvildarzadeh, A., Shoja, J., Alijani, S. (2006). Molecular characterization of CSN3 alleles in Sarabi and Holstein using PCR-RFLP. Biotechnology 5(4): 495-500. https://doi.org/10.3923/biotech.2006.495.500
  49. Miluchova, M., Gabor, M., Candrak, J., Trakovicka, A., Candrakova, K. (2018). Association of HindIII-polymorphism in kappa-casein gene with milk, fat and protein yield in Holstein cattle. Acta Biochimica Polonica 65(3): 403-407. https://doi.org/10.18388/abp.2017_2313 PMid:30212591          
  50. Ng-Kwai-Hang, K.F., Zadworny, D., Hayes, J.F., Kuhnlein, U. (1991). Identification of k-casein genotype in Holstein sires: A comparison between analysis of milk samples from daughters and direct analysis of semen samples from sires by polymerase chain reaction. J Dairy Sci. 74, 2410-2415. https://doi.org/10.3168/jds.S0022-0302(91)78415-4
  51. Bonvillani, G.A., Di Renzo, A.M., Tiranti, N.I. (2000). Genetic polymorphism of milk protein loci in Argentinian Holstein cattle. Genet Mol Biol. 23 (4): 819-823. https://doi.org/10.1590/S1415-47572000000400019
  52. Zepeda-Batista, J.L., Alacron-Zuniga, B., Ruiz-Flores, A., Nunez-Dominguez, R., Ramirez-Valverde, R. (2015). Polymorphism of three milk protein genes in Mexican Jersey cattle. Electron J Biotechn. 18, 1-4. https://doi.org/10.1016/j.ejbt.2014.10.002
  53. Erhardt, G., Prinzenberg, E.M., Buchberger, J., Krick-Saleck, H., Krause, I., Miller, M. (1997). Bovine κ-casein G detection, occurrence, molecular genetic characterization, genotyping and coagulation properties. International Dairy Federation special issue 2, 328-329.          
  54. Caroli, A., Bolla, P., Budelli, E., Barbieri, G., Leone, P. (2000). Effect of κ-casein E allele on clotting aptitude of Italian Friesian milk. Zoot Nutr Anim. 26(3): 127-130.        
  55. Barroso, A., Dunner, S., Canon, J. (1998). Technical note: Detection of bovine kappa casein variants A, B, C and E by means of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). J Anim Sci. 76, 1535-1538. https://doi.org/10.2527/1998.7661535x PMid:9655572            
  56. Jann, O.C., Ibeagha-Awemu, E.M., Ozbeyaz, C., Zaragoza, P., Williams, J.L., Ajmone-Marsan, P., Lenstra, J.A., Moazami-Goudarzi, K., Erhardt, G. (2004). Geographic distribution of haplotype diversity at the bovine casein locus. Genet Sel Evol. 36, 243-257. https://doi.org/10.1186/1297-9686-36-2-243 PMid:15040901 PMCid:PMC2697188         
  57. Boettcher, P.J., Caroli, A., Stella, A., Chessa, S., Budelli, E., Canavesi, F., Ghiroldi, S., Pagnacco, G. (2004). Effects of casein haplotypes on production traits in Italian Holstein and Brown Swiss cattle. J Dairy Sci. 87, 4311-4317. https://doi.org/10.3168/jds.S0022-0302(04)73576-6
  58. Schlieben, S., Erhardt, G., Senft, B. (1991). Genotyping of bovine κ-casein (κ-CNA, κ-CNB, κ-CNC, κ-CNE) following DNA sequence amplification and direct sequencing of κ-CNE PCR product. Anim Genet. 22, 333-342. https://doi.org/10.1111/j.1365-2052.1991.tb00687.x PMid:1683188             
  59. Chen, S.Y., Costa, V., Azevedo, M., Baig, M., Malmakov, N., Luikart, G., Erhardt, G., Beja-Pereira, A. (2008). Short communication: New alleles of the bovine κ-casein gene revealed by resequencing and haplotype inference analysis. J Dairy Sci. 91, 3682-3686. https://doi.org/10.3168/jds.2008-1211 PMid:18765626 
  60. Comin, A., Cassandro, M., Chessa, S., Ojala, M., Dal Zotto, R., De Marchi, M., et al. (2008). Effects of composite β- and κ-casein genotypes on milk coagulation, quality, and yiel traits in Italian Holstein cows. J Dairy Sci. 91, 4022-4027. https://doi.org/10.3168/jds.2007-0546 PMid:18832228

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© 2020 Adamov N. This is an open-access article published under the terms of the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Conflict of Interest Statement

The authors have declared that no competing interests exist.

Citation Information

Macedonian Veterinary Review. Volume 43, Issue 1, Pages 45-54, e-ISSN 1857-7415, p-ISSN 1409-7621, DOI: 10.2478/macvetrev-2020-0013, 2020

The all content of the Macedonian Veterinary Review, except where otherwise noted, is licensed under a Creative Commons Attribution License.