Ramya P R, Beena V, Radhika G, Shynu M, Jayavardhanan K K
Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Mannuthy, Thrissur District, Kerala State, 680651, India.
Department of Veterinary Physiology, Kerala Veterinary and Animal Sciences University, Mannuthy, Kerala State, 680651, India.
Heliyon. 2022 Dec 24;8(12):e12578. doi: 10.1016/j.heliyon.2022.e12578. eCollection 2022 Dec.
Hypoxia-inducible factor (HIF)-1α is a transcription factor stabilized by hypoxia by inducing or suppressing the homeostatic regulatory gene expression, enabling tissues and cells to survive despite fluctuations in environmental circumstances. As the name implies, hypoxia-inducible factor-1 is secreted not only as a cellular response to hypoxia but also in heat stress and oxidative stress. The goal of this work was to determine the molecular characterisation of the gene coding region as well as the differences in HIF-1αprotein primary structure between Vechur cattle and other cattle breeds in the online databases. Total RNA was isolated from blood samples of 6 Vechur cattle using the trizol reagent method, and full-length c sequences of the α gene were sequenced. The base pair length of composite HIF-1cDNA of Vechur cattle and encoding ORFis 3956 bp and 2469 bp respectively. The 5'UTR was recognized to be 279 bp in length. The start codon was identified at nucleotide 280-282, the stop codon UGA at 2746-2748 bp and a 1208 bp 3'UTR which included a poly-A tail of 27 adenine residues. In a comparative analysis of the cDNA, point transitions causing guanine to adenine (GA) changes at 1211th and 2699th positions were noticed as a heterozygous condition in the whole 3956 bp sequence. These two SNVs in the coding regions were responsible for two amino acid changes in the deduced 823 amino acid sequence. Since the predicted amino acid arginine had been replaced with lysine at 311th and 807th positions, it showed 99.76 percent sequence identity with . The phylogenetic tree revealed that the HIF-1α protein of Vechur cattle had a lesser evolutionary distance from the same gene of related species emphasising the highly conserved nature of this particular protein. This structural variation observed in the present study should be evaluated on a larger population to assess its functional relevance for thermo-tolerance.
缺氧诱导因子（HIF）-1α是一种转录因子，通过诱导或抑制稳态调节基因表达在缺氧状态下被稳定，使组织和细胞能够在环境条件波动的情况下存活。顾名思义，缺氧诱导因子-1不仅作为细胞对缺氧的反应而分泌，在热应激和氧化应激中也会分泌。这项工作的目的是在在线数据库中确定Vechur牛与其他牛品种之间编码区基因的分子特征以及HIF-1α蛋白一级结构的差异。使用trizol试剂法从6头Vechur牛的血液样本中分离总RNA，并对α基因的全长c序列进行测序。Vechur牛复合HIF-1 cDNA的碱基对长度和编码的开放阅读框分别为3956 bp和2469 bp。5'非翻译区（UTR）长度被认定为279 bp。起始密码子在核苷酸280 - 282处被识别，终止密码子UGA在2746 - 2748 bp处，还有一个1208 bp的3'UTR，其中包括一个由27个腺嘌呤残基组成的多聚腺苷尾。在cDNA的比较分析中，在整个3956 bp序列中，第1211位和第2699位发生了导致鸟嘌呤到腺嘌呤（GA）变化的点突变，呈现杂合状态。编码区的这两个单核苷酸变异（SNV）导致推导的823个氨基酸序列中有两个氨基酸发生变化。由于在第311位和第807位预测的氨基酸精氨酸被赖氨酸取代，它与……显示出99.76%的序列同一性。系统发育树显示，Vechur牛的HIF-1α蛋白与相关物种同一基因的进化距离较小，强调了这种特定蛋白的高度保守性。本研究中观察到的这种结构变异应在更大的群体中进行评估，以评估其对耐热性的功能相关性。
