牦牛SND1基因特征序列分析及其蛋白在乳腺的表达

发布时间：2018-05-09 05:50

本文选题：牦牛 + SND　；参考：《兽类学报》2017年02期

【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,分析其生物学特性,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,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3 294 bp,含有2 733 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%;系统进化树表明牦牛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。
[Abstract]:The purpose of this study was to clone the lactation related gene SND1 of yak, analyze its biological characteristics and study its expression in mammary gland. The primary mammary gland epithelial cells were obtained by trypsin digestion. The primary mammary gland epithelial cells were purified for 3 generations. The SND1 gene was amplified and cloned by RT-PCR, sequenced and spliced, and the characteristics of SND1 gene were analyzed by using the related bioinformatics software. Immunohistochemistry and immunofluorescence techniques were used to localize yak SND1 gene encoding protein. The results were as follows: the whole sequence of yak SND1 gene was 3 294 BP, containing 2 733 BP ORF, which contained 20 kinds of amino acid. SND1 gene encoding protein was nonsecretory protein, non transmembrane protein, homology analysis showed that yak SND1 gene, bison and domestic cattle, The homology of Tibetan antelope, goat, pig, wild camel, horse, chimpanzee, human and Rattus norvegicus is 990.98 / 995 / 91and 91/ 91/ 91g / 90.The phylogenetic tree indicates that the evolutionary level between yak and bison and domestic cattle is close to that between yak and bison and domestic cattle, and is far from that of human and mouse. The results of immunohistochemical staining showed that SND1 protein was highly expressed in secretory epithelial cells (breast epithelial cells) and ductal epithelial cells, and weakly expressed in myoepithelial cells. Immunofluorescence showed that the SND1 protein was overexpressed in the nucleus of mammary epithelial cells and weakly expressed in the cytoplasm. These results provide a basis for further study on the regulation of lactation function of yak by SND1, and also provide references for the study of alpine mammals.
【作者单位】：甘肃农业大学动物医学院;
【基金】：国家自然科学基金(31472244) 甘肃省生物技术专项(GNSW-2013-23)
【分类号】：S823.85

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