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维丘尔牛Toll样受体基因启动子序列的特征分析

Characterization of promoter sequence of toll-like receptor genes in Vechur cattle.

作者信息

Lakshmi R, Jayavardhanan K K, Aravindakshan T V

机构信息

Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Thrissur, Kerala, India.

Centre for Advanced Studies in Animal Genetics and Breeding, College of Veterinary and Animal Sciences, Thrissur, Kerala, India.

出版信息

Vet World. 2016 Jun;9(6):626-32. doi: 10.14202/vetworld.2016.626-632. Epub 2016 Jun 21.

Abstract

AIM

To analyze the promoter sequence of toll-like receptor (TLR) genes in Vechur cattle, an indigenous breed of Kerala with the sequence of Bos taurus and access the differences that could be attributed to innate immune responses against bovine mastitis.

MATERIALS AND METHODS

Blood samples were collected from Jugular vein of Vechur cattle, maintained at Vechur cattle conservation center of Kerala Veterinary and Animal Sciences University, using an acid-citrate-dextrose anticoagulant. The genomic DNA was extracted, and polymerase chain reaction was carried out to amplify the promoter region of TLRs. The amplified product of TLR2, 4, and 9 promoter regions was sequenced by Sanger enzymatic DNA sequencing technique.

RESULTS

The sequence of promoter region of TLR2 of Vechur cattle with the B. taurus sequence present in GenBank showed 98% similarity and revealed variants for four sequence motifs. The sequence of the promoter region of TLR4 of Vechur cattle revealed 99% similarity with that of B. taurus sequence but not reveals significant variant in motifregions. However, two heterozygous loci were observed from the chromatogram. Promoter sequence of TLR9 gene also showed 99% similarity to B. taurus sequence and revealed variants for four sequence motifs.

CONCLUSION

The results of this study indicate that significant variation in the promoter of TLR2 and 9 genes in Vechur cattle breed and may potentially link the influence the innate immunity response against mastitis diseases.

摘要

目的

分析维丘牛(一种喀拉拉邦本土品种,其序列与普通牛相同)中 toll 样受体(TLR)基因的启动子序列,并找出可能归因于针对牛乳腺炎的先天免疫反应的差异。

材料与方法

从喀拉拉邦兽医与动物科学大学维丘牛保护中心饲养的维丘牛颈静脉采集血样,使用柠檬酸葡萄糖抗凝剂。提取基因组 DNA,并进行聚合酶链反应以扩增 TLRs 的启动子区域。通过桑格酶促 DNA 测序技术对 TLR2、4 和 9 启动子区域的扩增产物进行测序。

结果

维丘牛 TLR2 启动子区域序列与 GenBank 中存在的普通牛序列显示出 98%的相似性,并揭示了四个序列基序的变体。维丘牛 TLR4 启动子区域序列与普通牛序列显示出 99%的相似性,但在基序区域未发现明显变体。然而,从色谱图中观察到两个杂合位点。TLR9 基因的启动子序列与普通牛序列也显示出 99%的相似性,并揭示了四个序列基序的变体。

结论

本研究结果表明,维丘牛品种中 TLR2 和 9 基因启动子存在显著变异,可能与影响针对乳腺炎疾病的先天免疫反应有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9d/4937055/38db44bc04dd/VetWorld-9-626-g001.jpg

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