Department of Animal Genetics and Breeding, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Mohanpur, Nadia, West Bengal, 741252, India.
Division of Livestock and Fishery Management, ICAR-Research Complex for Eastern Region, Patna, Bihar, 800014, India.
Cell Stress Chaperones. 2019 Nov;24(6):1187-1195. doi: 10.1007/s12192-019-01042-7. Epub 2019 Oct 22.
This study was aimed to genetic profiling of heat shock protein 70 (Hsp70) gene in Murrah buffalo investigating 50 unrelated adult animals at ICAR-Research Complex for Eastern Region, Patna (India) in winter, spring, and summer. PCR ready genomic DNA samples and season-wise total RNA samples were prepared. The PCR products of Hsp70 eluted from agarose gel were sequenced and analyzed. The first-strand cDNA was synthesized and concentration was equalized to 25 ng/μl. Expression kinetics of mRNA transcripts in different seasons was studied using Brilliant SYBR Green QPCR technique and the data retrieved was analyzed by least-squares ANOVA. DNA sequencing by primer walking revealed four allelic variants of Hsp70 gene. Alignment study revealed one substitution in 5'UTR, six substitutions in coding region, and one addition in 3'UTR. The highest percent identity and negligible phylogenetic distance were found among the alleles and reference bovine sequences. The relative mRNA expression was significantly higher in summer when THI ≥ 84 than the spring and winter; fold change increased by 4.5 times in summer than the spring whereas found nearly half in winter. These findings can be useful for heat stress management in buffaloes and help in understanding the mechanism of thermo-regulation well.
本研究旨在对印度帕特纳农业研究理事会东部区域研究中心的 50 头成年无亲缘关系的摩拉水牛的热休克蛋白 70（Hsp70）基因进行遗传分析，分别在冬季、春季和夏季进行。准备好 PCR 用基因组 DNA 样本和季节性总 RNA 样本。从琼脂糖凝胶中洗脱 Hsp70 的 PCR 产物进行测序和分析。用 Brilliant SYBR Green QPCR 技术研究不同季节的 mRNA 转录本的表达动力学，并通过最小二乘法方差分析对获取的数据进行分析。引物漫步 DNA 测序揭示了 Hsp70 基因的四个等位基因变异体。比对研究显示，在 5'UTR 中有一个取代，在编码区中有六个取代，在 3'UTR 中有一个添加。等位基因和参考牛序列之间的同一性最高，遗传距离最小。THI≥84 时，夏季的相对 mRNA 表达明显高于春季和冬季；夏季的倍数变化比春季增加了 4.5 倍，而冬季则几乎减少了一半。这些发现可用于水牛的热应激管理，并有助于深入了解体温调节机制。
