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通过聚合酶链反应-限制性片段长度多态性、聚合酶链反应-单链构象多态性检测Toll样受体4基因的遗传多态性,以与患乳房炎的奶牛建立关联。

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.

作者信息

Gupta Pooja H, Patel Nirmal A, Rank D N, Joshi C G

机构信息

Department of Biochemistry, B. A. College of Agriculture, Anand Agriculture University, Anand, Gujarat, India.

Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Anand, Gujarat, India.

出版信息

Vet World. 2015 May;8(5):615-20. doi: 10.14202/vetworld.2015.615-620. Epub 2015 May 15.

Abstract

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 χ(2) 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.

摘要

目的

尝试研究牛DNA中Toll样受体4(TLR4)基因多态性与患乳腺炎奶牛的相关性。

材料与方法

在本研究中,分别从坎克雷吉牛(22头)和三元杂交牛(24头)中,通过聚合酶链反应(PCR)扩增出TLR4基因的两个片段,即长度为316 bp的T4CRBR1和382 bp的T4CRBR2。第一个位点通过单链构象多态性检测两个群体中的遗传多态性,第二个位点通过用限制性内切酶Alu I消化片段来检测。

结果

结果表明,在两个群体中均发现了两个位点的两个等位基因(A和B),多态性信息含量值表明这些基因具有高度多态性。χ²检验的统计结果表明,两个群体中的两个多态性位点符合哈迪-温伯格平衡(p<0.05)。同时,TLR4基因多态性对体细胞评分(SCS)的影响表明,T4CRBR1中具有等位基因a的牛的SCS低于等位基因B的牛(p<0.05)。因此,等位基因A可能在奶牛抗乳腺炎中起重要作用。

结论

牛乳腺炎性状与TLR4基因多态性之间的关系表明,牛TLR4基因可能在抗乳腺炎中起重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f31e/4774722/55ee63f376fd/VetWorld-8-615-g001.jpg

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