Veterinary World

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Research (Published online: 15-05-2015)

10. Genetic polymorphism of toll-like receptors 4 gene by polymerase chain reaction-restriction fragment length polymorphisms, polymerase chain reaction-single-strand conformational polymorphism to correlate with mastitic cows - Pooja H. Gupta, Nirmal A. Patel, D. N. Rank and C. G. Joshi

Veterinary World, 8(5): 615-620



   doi: 10.14202/vetworld.2015.615-620


Pooja H. Gupta: Department of Biochemistry, B. A. College of Agriculture, Anand Agriculture University, Anand, Gujarat, India;

Nirmal A. Patel: Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Anand, Gujarat,


D. N. Rank: Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Anand, Gujarat,


C. G. Joshi: Department of Animal Biotechnology, College of Veterinary Science and Animal Husbandry, Anand, Gujarat,



Received: 20-12-2014, Revised: 05-04-2015, Accepted: 15-04-2015, Published online: 15-05-2015


Corresponding author: Pooja H. Gupta, e-mail:

Citation: Gupta PH, Patel NA, Rank DN, Joshi CG (2015) Genetic polymorphism of toll-like receptors 4 gene by polymerase chain reaction-restriction fragment length polymorphisms, polymerase chain reaction-single-strand conformational polymorphism to correlate with mastitic cows, Veterinary World 8(5):615-620.

Aim: An attempt has been made to study the toll-like receptors 4 (TLR4) gene polymorphism from cattle DNA to correlate with mastitis cows.

Materials and Methods: In present investigation, two fragments of TLR4 gene named T4CRBR1 and T4CRBR2 of a 316 bp and 382 bp were amplified by polymerase chain reaction (PCR), respectively from Kankrej (22) and Triple cross (24) cattle. The genetic polymorphisms in the two populations were detected by a single-strand conformational polymorphism in the first locus and by digesting the fragments with restriction endonuclease Alu I in the second one.

Results: Results showed that both alleles (A and B) of two loci were found in all the two populations and the value of polymorphism information content indicated that these were highly polymorphic. Statistical results of χ2 test indicated that two polymorphism sites in the two populations fit with Hardy–Weinberg equilibrium (p˂0.05). Meanwhile, the effect of polymorphism of TLR4 gene on the somatic cell score (SCS) indicated the cattle with allele a in T4CRBR1 showed lower SCS than that of allele B (p<0.05). Thus, the allele A might play an important role in mastitis resistance in cows.

Conclusion: The relationship between the bovine mastitis trait and the polymorphism of TLR4 gene indicated that the bovine TLR4 gene may play an important role in mastitis resistance.

Keywords: cattle, mastitis, restriction fragment length polymorphisms, single-strand conformational polymorphism, somatic cell score, toll-like receptors 4 gene.

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