Eun Kiyoung, Hwang Seon-Ung, Jeon Hye-Min, Hyun Sang-Hwan, Kim Hyunggee
a Department of Biotechnology, School of Life Sciences and Biotechnology , Korea University , Seoul , Republic of Korea.
b Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University , Cheongju , Chungbuk , Republic of Korea.
Anim Biotechnol. 2016;27(2):126-32. doi: 10.1080/10495398.2015.1126719.
Comparing the coding and regulatory sequences of genes in different species provides information on whether proteins translated from genes have conserved functions or gene expressions are regulated by analogical mechanisms. Herein, we compared the coding and regulatory sequences of glial fibrillary acidic protein (GFAP) from humans, mice, and pigs. The GFAP gene encodes a class III intermediate filament protein expressed specifically in astrocytes of the central nervous system. On comparing the mRNA, regulatory region (promoter), and protein sequences of GFAP gene in silico, we found that GFAP mRNA 3'-untranslated region (3'-UTR), promoter, and amino acid sequences showed higher similarities between humans and pigs than between humans and mice. In addition, the promoter-luciferase reporter gene assay revealed that the pig GFAP promoter functioned in human astrocytes. Notably, the 1.8-kb promoter fragment upstream from transcription initiation site showed strongest transcriptional activity compared to 5.2-kb DNA fragment or other regions of GFAP promoter. We also found that pig GFAP mRNA and promoter activity increased in pig fibroblasts by human IL-1β treatment. Taken together, these results suggest that the regulatory mechanisms and functions of pig genes might be more similar to those of humans than mice, indicating that pigs, particularly miniature pigs, are a useful model for studying human biological and pathological events.
比较不同物种中基因的编码序列和调控序列，可以提供有关从基因翻译而来的蛋白质是否具有保守功能或基因表达是否受类似机制调控的信息。在此，我们比较了人类、小鼠和猪的胶质纤维酸性蛋白（GFAP）的编码序列和调控序列。GFAP基因编码一种III类中间丝蛋白，特异性表达于中枢神经系统的星形胶质细胞中。通过在计算机上比较GFAP基因的mRNA、调控区域（启动子）和蛋白质序列，我们发现GFAP mRNA的3'非翻译区（3'-UTR）、启动子和氨基酸序列在人类和猪之间的相似性高于人类和小鼠之间的相似性。此外，启动子-荧光素酶报告基因检测显示猪GFAP启动子在人类星形胶质细胞中起作用。值得注意的是，与5.2-kb DNA片段或GFAP启动子的其他区域相比，转录起始位点上游1.8-kb的启动子片段显示出最强的转录活性。我们还发现，通过人IL-1β处理，猪成纤维细胞中猪GFAP mRNA和启动子活性增加。综上所述，这些结果表明猪基因的调控机制和功能可能比小鼠更类似于人类，这表明猪，特别是小型猪，是研究人类生物学和病理学事件的有用模型。
