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.
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