OJAS  Vol.3 No.1 , January 2013
Bovine β-casein: Detection of two single nucleotide polymorphisms by bidirectional allele specific polymerase chain reaction (BAS-PCR) and monitoring of their variation
Abstract: Due to the functional importance of bovine milk protein polymorphisms, their correct discrimination is of great interest both from a scientific and practical point of view. Nowadays a large number of commercial platforms are available for semiautomated or fully automated SNP geno-typing. However, in some cases the use of simple and rather cheap methods is an effective tool to be implemented within one’s own laboratory for the routine analysis of a specific SNP. The present paper describes two simple tests based on the bidirectional allele-specific polymerase chain reaction (BAS-PCR) developed for the identification of β-casein (CSN2) B and I genetic variants. The practical application of the two methods on a panel of 84 Italian Brown bulls and 100 Italian Friesian cows is also discussed, including the biological significance of the two genetic variants and the importance of taking their occurrence into account when linkage analyses are performed on milk functional properties. A combined system for analysing milk protein variants by isoelectrofocusing (IEF) and the BAS-PCR assay developed for CSN2*I is described.
Cite this paper: Chessa, S. , Bulgari, O. , Rossoni, A. , Ceriotti, G. and Caroli, A. (2013) Bovine β-casein: Detection of two single nucleotide polymorphisms by bidirectional allele specific polymerase chain reaction (BAS-PCR) and monitoring of their variation. Open Journal of Animal Sciences, 3, 36-41. doi: 10.4236/ojas.2013.31005.

[1]   Caroli, A.M., Chessa, S. and Erhardt, G.J. (2009) Milk protein genetic variation in cattle: Impact on animal breeding and human nutrition. Journal of Dairy Science, 92, 5335-5352. doi:10.3168/jds.2009-2461

[2]   Farrell Jr, H.M., Jimenez-Flores, R., Bleck, G.T., Brown, E.M., Butler, J.E., Creamer, L.K., et al. (2004) Nomen-clature of the proteins of cows’ milk—Sixth revision. Journal of Dairy Science, 87, 1641-1674. doi:10.3168/jds.S0022-0302(04)73319-6

[3]   Formaggioni, P., Summer, A., Malacarne, M. and Mariani, P. (1999) Milk protein polymorphism: Detection and diffusion of the genetic variants in Bos genus. Annali della Facoltà di Medicina Veterinaria Università di Parma, 29, 127-165.

[4]   Jann, O., Ceriotti, G., Caroli, A. and Erhardt, G. (2002) A new variant in exon VII of bovine β-casein gene (CSN2) and its distribution among European cattle breeds. Journal of Animal Breeding and Genetics, 119, 65-68. doi:10.1046/j.1439-0388.2002.00318.x

[5]   Visker, M.H.P.W., Dibbits, B.W., Kinders S.M., van Valenberg, H.J.F., van Arendonk, J.A.M. and Bovenhuis H. (2010) Association of bovine β-casein protein variant I with milk production and milk protein composition. Animal Genetics, 42, 212-218. doi:10.1111/j.1365-2052.2010.02106.x

[6]   Aschaffenburg, R. (1966) Modified procedure of starch gel electrophoresis for β-casein phenotyping. Journal of Dairy Science, 49, 1284. doi:10.3168/jds.S0022-0302(66)88073-6

[7]   Krause, I., Buchberger, J., Weis, G. and Klostermeyer, H. (1988) Screening methods for genetic variants of milk proteins. In: Barth, C.A. and Schlimme, E. Eds., Milk Proteins: Nutritional, Clinical, Functional and Technological Aspects, Steinkopff Verlag, Darmstadt, 171-173.

[8]   Bech, A.-M. and Kristiansen, K.R. (1990) Milk protein polymorphism in Danish dairy cattle and the influence of genetic variants on milk yield. Journal of Dairy Research, 57, 53-62. doi:10.1017/S0022029900026601

[9]   Erhardt, G., Juszczak, J., Panicke, L. and Krick-Saleck, H. (1998) Genetic polymorphism of milk proteins in Polish Red Cattle: A new genetic variant of β-lactoglobulin. Journal of Animal Breeding and Genetics, 115, 63-71. doi:10.1111/j.1439-0388.1998.tb00328.x

[10]   Bonfatti, V., Grigoletto, L., Cecchinato, A., Gallo, L. and Carnier, P. (2008) Validation of a new reversed-phase high-performance liquid chromatography method for separation and quantification of bovine milk protein genetic variants. Journal of Chromatography A, 1195, 101-106. doi:10.1016/j.chroma.2008.04.075

[11]   Keating, A.F., Smith, T.J., Ross, R.P. and Cairns, M.T. (2006) A single nucleotide polymorphism in the bovine beta-casein promoter region across different bovine breeds. Journal of Dairy Research, 73, 193-196. doi:10.1017/S0022029905001615

[12]   Damiani, G., Pilla, F., Leone, P. and Cacciò, S. (1992) Direct sequencing and bidirectional allele specific poly- merase chain reaction of the bovine beta-casein B variant. Animal Genetics, 23, 561-565. doi:10.1111/j.1365-2052.1992.tb00180.x

[13]   Lien, S., Alestrom, P., Hlungland, H. and Rogne, S. (1992) Detection of multiple beta-casein (CASB) alleles by amplification created restriction sites (ACRS). Animal Genetics, 23, 333-338. doi:10.1111/j.1365-2052.1992.tb00155.x

[14]   Barroso, A., Dunner, S. and Canon, J. (1999) A multiplex PCR-SSCP test to genotype bovine beta-casein alleles A1, A2, A3, B, and C. Animal Genetics, 30, 322-323. doi:10.1046/j.1365-2052.1999.00445-6.x

[15]   Chessa, S., Chiatti, F., Ceriotti, G., Caroli, A., Consolandi, C., Pagnacco, G. and Castiglioni, B. (2007) Development of a SNP genotyping microarray platform for the identification of bovine milk protein genetic polymorphisms. Journal of Dairy Science, 90, 451-464. doi:10.3168/jds.S0022-0302(07)72647-4

[16]   Pariset, L., Caroli, A., Chessa, S., Fontanesi, L., Russo, V., Bagnato, A., et al. (2009) Assessment of 29 candidate genes for milk traits in Italian dairy cattle. Italian Journal of Animal Science, 8, 226.

[17]   Rozen, S. and Skaletsky, H.J. (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz, S. and Misener, S., Eds., Bioinformatics Methods and Protocols: Methods in Molecular Biology, Humana Press, Totowa, 365-386.

[18]   Caroli, A., Chessa, S., Vivona, G., Bolla, P. and Pagnacco, G. (2003) Analysis of milk protein polymorphisms and casein haplotypes in Italian Friesian, Italian Brown Swiss and Reggiana cattle. Book of Abstract of the 54th Annual Meeting of the European Association for Animal Produc- tion, Roma, 31 August-3 September 2003, 97.

[19]   Xie, X. and Ott, J. (1993) Testing linkage disequilibrium between a disease gene and marker loci. American Journal of Human Genetics, 53, 1107.

[20]   Boettcher, P.J., Caroli, A., Stella, A., Chessa, S., Budelli, E., Canavesi, F., et al. (2004) Effects of casein haplotypes on production traits in Italian Holstein and Brown Cattle. Journal of Dairy Science, 87, 4311-4317. doi:10.3168/jds.S0022-0302(04)73576-6

[21]   Chessa, S., Rignanese, D., Conte, G., Severgnini, M., Ceriotti, G., Caroli, A., Pagnacco, G., Castiglioni, B. (2008) Application of a microarray technology to SNPs detection within genes involved in milk production. Chromosome Research, 16, 1041-1042.

[22]   Mariani, P. (1997) Contenuto di caseina, varianti genetiche ed attitudine tecnologico-casearia del latte delle vacche di razza bruna nella produzione del formaggio grana. La razza Bruna italiana, 2, 8-14.

[23]   Mariani, P. (1999) Polimorfismo delle proteine ed attitudine tecnologico-casearia del latte. La razza Bruna italiana, 1, 1-7.

[24]   Hayes, B.J., Bowman, P.J., Chamberlain, A.J. and Goddard, M.E. (2009) Invited review: Genomic selection in dairy cattle: Progress and challenges. Journal of. Dairy Science, 92, 433-443. doi:10.3168/jds.2008-1646

[25]   Heck, J.M.L., Schennink, A., van Valenberg, H.J.F., Bovenhuis, H., Visker, M.H.P.W., van Arendonk, J.A.M., et al. (2009) Effects of milk protein variants on the protein composition of bovine milk. Journal of Dairy Science, 92, 1192-1202. doi:10.3168/jds.2008-1208