RFLP Analysis of Beta‐Lactoglobulin Gene in Swamp and Murrah Buffaloes Using a Single Restriction Enzyme

Document Type : Short Communication


1 Research Center for Bioscience in Animal Production, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand

2 Murrah Farm, 99 / 14 Moo 12 Nongsoiting, Plangyao, Chacheongsao, 24000, Thailand


An attempt has been made to analyze the distribution of the beta-lactoglobulin genotype in swamp buffaloes and Murrah buffaloes utilizing polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Blood samples were taken from 50 swamp buffaloes and 50 Murrah buffaloes. The DNA was extracted by the phenol-chloroform method. The PCR-RFLP was performed using beta-lactoglobulin gene primers and a single restriction enzyme, Hae III. The enzyme digested products were separated by electrophoresis on 2.5%agarose gel. All the 100 DNA samples of swamp and Murrah buffaloes resulted in 398 bp product on amplifying beta-lactoglobulin gene. Those PCR products (398 bp fragment) were digested with Hae III produced only one type of restriction pattern yielding five fragments of 113, 99, 89, 73 and 24 bp. One hundred DNA samples of swamp and Murrah buffalo were examined in this study and revealed no polymorphism at the beta-lactoglobulin gene locus.


Badola S., Bhattacharya T.K., Biswas T.K., Shivakumar B.M., Kumar P., Mistra S.S. and Sharma A. (2003). Bubaline beta-lactoglobulin gene polymorphism: A contrast from its bovine counterpart. Buffalo J. 19(4), 291-298.
Badola S., Bhattacharya T.K., Biswas T.K., Shivakumar B.M., Kumar P. and Sharma A. (2004). A Comparison on polymorphism of beta-lactoglobulin gene in Bos indicus, Bos taurus and Indicine x taurine crossbred cattle. Asian-Aust. J. Anim. Sci. 17(6), 733-736.
Del Lama S.N. and Zago M.A. (1996). Identification of the K-casein and B-lactoglobulin genotypes in Brazilian Bos indicus and Bubalus bubalis populations. Brazilian J. Genet. 19(1), 73-77.
Karimi K., Belgi Nasiri M.T., Fayyazi J., Mirzadeh K.H. and Roushanfekr H. (2009). Allele and genotype frequencies of B-lactoglobulin gene in Iranian Najdi cattle and buffalo populations using PCR-RFLP. African J. Biotech. 8(15), 3654-3657.
Meignanalakshmi A., Mahalinga Nainar A. and Nachimuthu K. (2001). Identification of genetic polymorphism of beta-lactoglobulin gene locus in Red Sindhi cows by PCR-RFLP analysis. Int. J. Anim. Sci. 16(2), 223-226.
Meignanalakshmi S. and Mahalinga Nainar A. (2009). PCR-RFLP analysis of beta-lactoglobulin gene in Murrah buffaloes Tamilnadu J. Vet. Anim. Sci. 5(5), 194-197.
Ng-Kwai-Hang K.F., Hayes J.F., Moxley J.E. and Monardes H.G. (1984). Association of genetic variants of casein and milk serum protein with milk fat and protein production by dairy cattle. J. Dairy Sci. 67, 835-840.
Ng-Kwai-Hang K.F., Zadworny D., Hayes J.F. and KuhnleinU.(1992). Identification of k-casein genotype in Holsteinsires: 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.
Pipalia D.L., Ladani D.D., Brahmshtri B.P., Rank D.N., Joshi C.G., Vataliya P.H. and Solanki J.V. (2001). Kappa-casein genotyping of Indian buffalo breeds using PCR-RFLP. Buffalo J. 2, 195-202.
Sambrook J., Fritsch E.F. and Maniatis T. (1989). Molecular Cloning: A Laboratory Manual. 2nd Ed.ColdSpringHarbor.