Polymorphism study of TLR4 gene in crossbred cattle of Kerala

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Abstract

The present study was conducted to identify single nucleotide variations in exon 3 region of the bovine Toll-Like Receptor gene (TLR4) associated with somatic cell score (SCS) in crossbred cattle of Kerala using polymerase chain reaction-single stranded conformational polymorphisms (PCR-SSCP) technique. Blood samples were collected from 200 lactating crossbred cattle in different farms of Kerala Veterinary and Animal Sciences University (KVASU) and field centres of ICAR- FPT scheme, KVASU. Genomic DNA was isolated and polymorphisms were identified by SSCP analysis. The 231 bp fragment of exon 3 in TLR4 gene was found to polymorphic and two genotypes viz. CC and CD were obtained with frequencies 77 per cent and 23 per cent respectively. The frequencies of C and D allele in crossbred were found to be 0.88 and 0.12. On sequencing, C to T transition had led to one non-synonymous change at 2021th position of ORF. Cattle with CC genotype were associated with significantly lower SCS (p≤0.05) as compared to CD genotype. The association between TLR4 polymorphism and SCS described in this study suggests the possible role of SNP during mastitis infection. Thus, could be employed as a useful marker for genetic selection to promote mastitis resistance in dairy cattle.
喀拉拉邦杂交牛TLR4基因多态性研究
本研究采用聚合酶链反应-单链构象多态性(PCR-SSCP)技术,对喀拉拉邦杂交牛体细胞评分(SCS)相关的toll样受体基因(TLR4)外显子3区单核苷酸变异进行了鉴定。从喀拉拉邦兽医和动物科学大学(KVASU)的不同农场和KVASU ICAR- FPT计划的现场中心收集了200头泌乳杂交牛的血液样本。分离基因组DNA,通过SSCP分析鉴定多态性。发现TLR4基因外显子3 231 bp片段多态性,获得CC和CD两种基因型,频率分别为77%和23%。C和D等位基因的频率分别为0.88和0.12。在测序上,C到T的转变导致ORF的第2021位发生了一个非同义变化。与CD基因型相比,CC基因型牛的SCS显著降低(p≤0.05)。本研究描述的TLR4多态性与SCS之间的关联提示SNP在乳腺炎感染中的可能作用。因此,可以作为一种有用的遗传选择标记来促进奶牛对乳腺炎的抗性。
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