Marker Assisted Molecular Investigation of Kappa-Casein Gene in Bos Indicus Sindhi Genetic Group Using HINFI Restriction Enzyme
Background: Present research aimed to determine molecular genotype of kappa-casein gene in female Red Sindhi cattle. This gene has great influence on the technologically advanced milk properties.
Methods: Blood specimens (n=50) from females of this cattle breed at a well-managed farm in Sindh-Pakistan were collected and commercial kit was employed for DNA extraction. Genotype determination of κ-casein gene and alleles was done through PCR-RFLP technique by using primer; PCR products were digested upon HINFI restriction enzyme. The digested fragments were analyzed by electrophoresis on agarose gel using ethidium bromide to increase visibility. The bands were examined under ultra violet-light to study polymorphic locus on DNA fragments.
Results: Digestion upon HINFI restriction enzyme of 350bp fragment indicated three patterns. The 1st (homozygote genotype BB), 2nd (homozygote genotype AA) and 3rd (heterozygote genotype AB) patterns yielded major fragment(s) of 1) 266bp, 2) 134bp and 132bp and 3) 134bp, 132bp and 266bp, respectively. Each of the three patterns yielded one minor fragment of 84bp. The genotype frequency for homozygote AA and the allelic frequency of allele A were higher than the same for homozygote genotype BB and the allelic frequency of allele B, respectively.
Conclusions: An accurate profile of genetic make-up and alternate forms of κ-casein genes in Sindhi cattle is likely to help researchers, policy makers, immunologists, dietitians, neonatologists, community physicians and managerial as well as production level officials to exploit it to full potential.
2. Stelwagen, K, E Carpenter, B Haigh, A Hodgkinson, TT Wheeler. Immune components of bovine colostrum and milk 1. Dairy Science & Technology, 2009;87(13 Suppl):3-9. doi: 10.2527/jas.2008-1377.
3. Holmer-Jensen, J, T Karhu, LS Mortensen, SB Pedersen, K-H Herzig, K Hermansen. Differential effects of dietary protein sources on postprandial low-grade inflammation after a single high fat meal in obese non-diabetic subjects. Nutr J, 2011;10:115.
4. Lisson, M. Appearance of epitopes in bovine milk protein variants, their allergenicity and potential use in human nutrition. 2014; PhD dissertation, Department of Animal Breeding and Genetics, Justus‐Liebig‐University Gießen, Germay.
5. Ahmad L, Dogar MZH. Insecticidal exposure may be the cause of progression of immune disorders and inflammatory diseases through dys-regulation of cytokines. Int Arch BioMed Clin Res. 2016;2(3):18-27. doi:10.21276/iabcr.2016.2.3.4
6. Leghari, MF, ZA Nizamani, IH Leghari, TM Samo, A Sethar, F Samo. Hemato-biochemical and pathological studies on induced Pastuerella Multocida infection in rabbits. Sindh University Research Journal (SURJ), 2016;48(3):671-674.
7. Kiplagat, SK, MK Limo and IS Kosgey. Genetic improvement of livestock for milk production. In: Milk Production – Advanced Genetic Traits, Cellular Mechanism, Animal Management and Health Edited by Narongsak Chaiyabutr. ISBN 978-953-51-0766-8 doi:10.5772/2475:77-96.
8. Duifhuis-Rivera, T, C Lemus-Flores, MÁ Ayala-Valdovinos, DR Sánchez -Chiprés, J Galindo-García, K Mejía-Martínez, et al. Polymorphisms in beta and kappa-casein are not associated with milk production in two highly technified populations of Holstein cattle in Mexico. J Anim Plant Sc, 2014;24(5):1316-1321.
9. Gouda, EM, MKh. Galal, SA Abdelaziz. Genetic variants and allele frequencies of kappa casein in Egyptian cattle and buffalo using PCR-RFLP. J Agric. Sc, 2013;5(2):197-203.
10. Ferretti, L, P Leone, V Sgaramella.. Long range restriction analysis of the bovine casein genes. Nucleic Acids Res. 1990;18:6829-6833.
11. Cinar, MU, B Akyuz, K Arslan, EA Ilgar. Genotyping of the kappa-casein and beta-lactoglobulin genes in Anatolian water buffalo by PCR-RFLP. Int J Dairy Technol, 2016;69(2):308-311.
12. Khan, BB, Isani GB. Animal Husbandry. National Book Foundation, Islamabad, 2013;53-54.
13. Sahito, IA, JGM Sahito, IUK Wazir, GM Lochi. Analysis of the persistency of lactation in Red Sindhi cattle. Sci Int (Lahore), 2016;28(3):2607-2610.
14. Medrano, JF, E Aguilar-Cordova. Genotyping of bovine kappa casein loci following DNA sequencing amplification. J. Biotech. 1990;8:144-146.
15. Weir, BS, 1996. Genetic data analysis In: Methods for Discrete Population Genetic Data. 2nd ed. Sinauer Associates, Suderland.
16. Azevedo, ALS, CS Nascimento, RS Steinberg, MRS Carvalho, MGCD Peixoto1, RL Teodoro et al. Genetic polymorphism of the kappa-casein gene in Brazilian cattle. Genet Molec Res. 2008;7:623-630.
17. Haldane, JBS. An exact test for randomness of mating. J Genet,1954;52:631-635.
18. Miller, MP. Tools for population genetics analyses (TFPGA). A WINDOWS program for the analysis of allozyme and molecular population genetic data. 1997. http://www.marksgeneticsoftware.net/tfpga.htm
19. Patel, G, RK Patel, KJ Soni. Detection of κ-casein and β-lactoglobulin variants in Holstein Friesian cattle by PCR-RFLP assays. Haryana Veterinarian, 2007;46:48-51.
20. Gangaraj, DR, S Shetty, MG Govindaisah, CS Nagaraja, SM Byregowda, MR Jayashankar. Molecular characterization of kappa-casein gene in buffaloes. Sci Asia, 2008;34:435-439.
21. El-Rafey, GA, SF Darwish,. A PCR-RFLP assay to detect genetic variants of kappa-casein gene in cattle and buffalo. Arab J Biotech, 2007;11(1):11-18.
22. Sitkowska, B, W Neja, E Wiśniewska. Relations between Kappa-casein (CSN3) polymorphism and milk performance traits in heifer cows. J Central European Agric, 2008;9:641-644.
23. Dogru, U, M Ozdemir. Genotyping of kappa casein locus by PCR-RFLP in brown Swiss cattle breed. J Anim Vet Advances. 2009;8(4):779-781
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