本文選題：牦牛 + SND�。� 參考：《獸類學報》2017年02期

【摘要】：Staphylococcal nuclease and tudor domain containing 1(SND1,Tudor-SN)是一種參與基因調(diào)控的轉(zhuǎn)錄共激活因子蛋白,本研究意在克隆牦牛泌乳相關(guān)基因SND1,分析其生物學特性,研究其蛋白在乳腺的表達。采集牦牛泌乳期乳腺組織,胰蛋白酶消化法得到原代乳腺上皮細胞,純化到3代,采用RT-PCR擴增克隆SND1基因,測序并拼接,用相關(guān)生物信息軟件分析牦牛SND1基因特性;用免疫組織化學和免疫熒光技術(shù)對牦牛SND1基因編碼蛋白進行定位分析。獲得如下結(jié)果:牦牛SND1基因全序列為3 294 bp,含有2 733 bp的ORF,共包含20種氨基酸。SND1基因編碼蛋白為非分泌蛋白,非跨膜蛋白;同源性分析顯示,牦牛SND1基因與野牛、家牛、藏羚羊、山羊、豬、野駱駝、馬、黑猩猩、人、褐家鼠的同源性分別為99%、98%、96%、94%、91%、90%、90%、89%、89%、85%;系統(tǒng)進化樹表明牦牛與野牛和家牛的進化水平較近,與人和鼠的進化水平較遠。免疫組織化學染色結(jié)果顯示,SND1蛋白在分泌上皮細胞(乳腺上皮細胞)和導管上皮細胞呈陽性高表達,在肌上皮細胞呈弱表達。免疫熒光顯示,SND1蛋白在乳腺上皮細胞胞核高表達,胞質(zhì)弱表達。上述研究結(jié)果為進一步探究SND1對牦牛泌乳機能的調(diào)節(jié)提供了相關(guān)依據(jù),也為高寒哺乳動物的研究提供了參考資料。
[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.
【作者單位】： 甘肅農(nóng)業(yè)大學動物醫(yī)學院;
【基金】：國家自然科學基金(31472244) 甘肅省生物技術(shù)專項(GNSW-2013-23)
【分類號】：S823.85

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