ACTA THERIOLOGICA SINICA ›› 2017, Vol. 37 ›› Issue (2): 207-214.

Previous Articles    

The characteristics of yak SND1 gene and its expression in mammary gland

TIAN Kuili, PAN Yangyang, LI Yueqin, CUI Yan, YU Sijiu   

  1. College of Veterinary Medicine,Gansu Agricultural University
  • Online:2017-04-20 Published:2017-04-20

牦牛SND1基因特征序列分析及其蛋白在乳腺的表达

田奎丽 潘阳阳 李月琴 崔燕 余四九   

  1. 甘肃农业大学 动物医学院

Abstract:

Staphylococcal nuclease and tudor domain containing 1 (SND1, Tudor-SN), a transcriptional co-activator protein, is involved in the regulation of gene expression. The objective of this study was to analyze the biological characteristic of yak SND1, and to examine the expression of SND1 in mammary gland. Mammary samples from local yak were collected and primary mammary epithelial cells were obtained by trypsin digestion. The third generation of purified mammary epithelial cells was used to clone yak SND1 gene by RT-PCR. Bio-information software was used to analyze the characteristics of SND1, and immunohistochemistry and immunofluorescence were used to detect the location of protein encoded by SND1. The results showed that the coding region of Bos grunniens SND1 gene was 3294 bp in length, encoding 20 amino acids. The encoded protein of SND1 gene was predicted to be a type of non-secreted and non-transmembrane protein. The homology of SND1 gene between yak and Bison, Bos taurus, Pantholops hodgsonii, Capra hircus, Sus scrofa, Camelus ferus, Equus caballus, Pan paniscus, Homo sapiens, Rattus norvegicus were 99%, 98%, 96%, 94%, 91%, 90%, 90%, 89%, 89% , 85%, respectively. It was found that, in evolution, the SND1 gene of yak was closely related to those of Bison and, Bos taurus, and distantly related to that of Homo sapiens and Rattus norvegicus. In yak mammary gland, the SND1 had high positive expression in the secretory epithelial cells (mammary epithelial cell) and ductal epithelial cells, but weak expression in myo-epithelial cells. SND1 protein was highly expressed in the nuclei of mammary epithelial cells and weakly expressed in cytoplasm. In summary, our study could provide a theoretical basis for further study of yak lactation function, and also provide reference data for the mammals living in Qinghai-Tibet Plateau.
 

Key words: Yak, SND1, Mammary epithelial cells, Biological characteristic, Expression

摘要: Staphylococcal nuclease and tudor domain containing 1( SND1,Tudor-SN)是一种参与基因调控的转录共激活因子蛋白,本研究意在克隆牦牛泌乳相关基因SND1,分析其生物特性,研究其蛋白在乳腺的表达。采集牦牛泌乳期乳腺组织,胰蛋白酶消化法得到原代乳腺上皮细胞,纯化到3代, 采用RT-PCR扩增克隆SND1基因,测序并拼接,并用相关生物信息软件分析牦牛SND1基因特性;用免疫组织化学和免疫荧光技术对牦牛SND1基因编码蛋白进行定位分析。获得如下结果:牦牛SND1基因全序列为3294 bp,含有2733 bp的ORF,共包含20种氨基酸。SND1基因编码蛋白为非分泌蛋白,非跨膜蛋白;同源性分析显示,牦牛SND1基因与野牛、家牛、藏羚羊、山羊、猪、野骆驼、马、黑猩猩、人、褐家鼠的同源性分别为99%、98%、96%、94%、91%、90%、90%、89%、89%、85%;系统进化树表明与野牛和家牛的进化水平较近,与人和鼠的进化水平较远。免疫组织化学染色结果显示,SND1蛋白在分泌上皮细胞(乳腺上皮细胞)和导管上皮细胞呈阳性高表达,在肌上皮细胞呈弱表达。免疫荧光显示,SND1蛋白在乳腺上皮细胞胞核高表达,胞质弱表达。上述研究结果为进一步探究SND1对牦牛泌乳机能的调节提供了相关依据,也为高寒哺乳动物的研究提供了参考资料。

关键词: 牦牛, SND1, 乳腺上皮细胞, 生物学特性, 表达