Buffalo Genomics Laboratory, National Bureau of Animal Genetic Resources, Karnal, Haryana, India.
Anim Biotechnol. 2011 Jul-Sep;22(3):124-32. doi: 10.1080/10495398.2011.581558.
The present study was undertaken to characterize the structure of S100A8 gene and its promoter in water buffalo and yak. Sequence data of 2.067 kb, 2.071 kb, and 2.052 kb with respect to complete S100A8 gene including 5' flanking region was generated in river buffalo, swamp buffalo, and yak, respectively. BLAST analysis of coding DNA sequences (CDS) of S100A8 gene revealed 95% homology of buffalo sequence with cattle, 85% with pig and horse, 83% with dog, 72-73% with murines, and around 79% with primates and humans. Phylogenetic analysis of predicted CDS revealed distinct clustering of murines, primates, and domestic animals with bovines and bubalines forming a subcluster among farm animals. In silico translation of predicted CDS revealed a sequence of 89 amino acids with 7 amino acid changes between cattle and buffalo and 2 changes between cattle and yak. The search for Pfam family revealed the N-terminal calcium binding domain and the noncanonical EF hand domain in the carboxy terminus, with more variations being observed in the N-terminal domain among different species. Two amino acid changes observed in carboxy terminal EF hand domain resulted in altered secondary structure of yak S100A8 protein. Analysis of S100A8 gene promoter revealed 14 putative motifs for transcriptional factor binding sites. Two putative motifs viz. C/EBP and v-Myb were found to be absent in swamp buffalo as compared to river buffalo and cattle. Differences in the structure of S100A8 protein and the transcriptional factor binding sites identified in the present study need to be analyzed further for their functional significance in yak and swamp buffalo respectively.
本研究旨在对水牛和牦牛的 S100A8 基因及其启动子结构进行研究。从河流水牛、沼泽水牛和牦牛中分别获得了全长为 2.067kb、2.071kb 和 2.052kb 的 S100A8 基因及其 5'侧翼区序列。对 S100A8 基因编码 DNA 序列（CDS）的 BLAST 分析表明，水牛序列与牛的同源性为 95%，与猪和马的同源性为 85%，与狗的同源性为 83%，与鼠的同源性为 72-73%，与灵长类动物和人类的同源性约为 79%。预测 CDS 的系统发育分析表明，鼠、灵长类动物和家畜与牛和瘤牛形成一个亚群，在农场动物中形成一个聚类。预测 CDS 的体外翻译揭示了 89 个氨基酸的序列，牛和水牛之间有 7 个氨基酸变化，牛和牦牛之间有 2 个氨基酸变化。对 Pfam 家族的搜索揭示了 N 端钙结合域和羧基末端的非典型 EF 手域，在不同物种中，N 端结构域存在更多的变异。在羧基末端 EF 手域观察到的两个氨基酸变化导致了牦牛 S100A8 蛋白的二级结构发生改变。S100A8 基因启动子分析显示有 14 个潜在的转录因子结合位点的假定模体。与河流水牛和牛相比，沼泽水牛中发现有两个假定的模体，即 C/EBP 和 v-Myb 缺失。在本研究中鉴定的 S100A8 蛋白结构和转录因子结合位点的差异需要进一步分析，以了解它们在牦牛和沼泽水牛中的功能意义。
