藏羊白细胞介素-7基因的PCR扩增及生物信息学分析
发布时间：2018-09-17 10:24
【摘要】：为了获得藏羊白细胞介素-7(interleukin-7,IL-7)基因序列,并研究其序列特征及编码蛋白的结构和功能,本试验采用RT-PCR方法,从藏羊脾脏中扩增了IL-7基因,应用DNAStar软件分析该基因的核苷酸和氨基酸序列,与经BLAST比对后的参考序列进行同源性比对,并构建系统进化树,同时利用DNAStar软件和在线服务器预测该基因编码蛋白的二级结构、三级结构、亲水性、信号肽和蛋白翻译后修饰位点。结果表明,藏羊IL-7基因长度为531bp(含终止密码子),编码176个氨基酸。藏羊IL-7基因与绵羊、山羊、藏羚羊、水牛、瘤牛、美洲草原野牛、黄牛和牦牛的IL-7基因核苷酸序列同源性在97.2%~99.8%之间,氨基酸序列同源性在94.9%~99.4%之间,藏羊与绵羊的亲缘关系最近。蛋白结构预测结果表明,IL-7蛋白主要由α螺旋组成,是一种亲水性和分泌型蛋白。该蛋白含有6种蛋白质翻译后修饰位点,包括1个N-豆蔻酰化位点、1个酰胺化位点、1个cAMP和cGMP依赖性蛋白激酶磷酸化位点、3个N-糖基化位点、4个蛋白激酶C磷酸化位点、6个酪蛋白激酶Ⅱ磷酸化位点。本研究结果可为藏羊IL-7基因的进一步研究与临床应用提供参考。
[Abstract]:In order to obtain the sequence of interleukin-7 (interleukin-7,IL-7) gene of Tibetan sheep and study its sequence characteristics and the structure and function of encoded protein, the IL-7 gene was amplified from the spleen of Tibetan sheep by RT-PCR method. The nucleotide and amino acid sequences of the gene were analyzed by DNAStar software, and compared with the reference sequence after BLAST alignment. The phylogenetic tree was constructed. The secondary structure of the gene encoded protein was predicted by DNAStar software and online server. Tertiary structure, hydrophilicity, signal peptide and protein posttranslational modification site. The results showed that the length of IL-7 gene of Tibetan sheep was 531bp, encoding 176 amino acids. The nucleotide sequence homology of IL-7 gene of Tibetan sheep and sheep, goat, Tibetan antelope, buffalo, cattle, buffalo, yellow cattle and yak was 97.2% and 99.8% respectively, and the homology of amino acid sequence between Tibetan sheep and sheep was 99.4%. The relationship between Tibetan sheep and sheep was the most close. The predicted structure of IL-7 protein is mainly composed of 伪 helix and is a hydrophilic and secretory protein. The protein contains six post-translational modification sites. There were one N-nutmeg site, one amidation site, one cAMP and cGMP dependent protein kinase phosphorylation site, three N-glycosylation sites, four protein kinase C phosphorylation sites and six casein kinase 鈪，
本文编号：2245553
[Abstract]:In order to obtain the sequence of interleukin-7 (interleukin-7,IL-7) gene of Tibetan sheep and study its sequence characteristics and the structure and function of encoded protein, the IL-7 gene was amplified from the spleen of Tibetan sheep by RT-PCR method. The nucleotide and amino acid sequences of the gene were analyzed by DNAStar software, and compared with the reference sequence after BLAST alignment. The phylogenetic tree was constructed. The secondary structure of the gene encoded protein was predicted by DNAStar software and online server. Tertiary structure, hydrophilicity, signal peptide and protein posttranslational modification site. The results showed that the length of IL-7 gene of Tibetan sheep was 531bp, encoding 176 amino acids. The nucleotide sequence homology of IL-7 gene of Tibetan sheep and sheep, goat, Tibetan antelope, buffalo, cattle, buffalo, yellow cattle and yak was 97.2% and 99.8% respectively, and the homology of amino acid sequence between Tibetan sheep and sheep was 99.4%. The relationship between Tibetan sheep and sheep was the most close. The predicted structure of IL-7 protein is mainly composed of 伪 helix and is a hydrophilic and secretory protein. The protein contains six post-translational modification sites. There were one N-nutmeg site, one amidation site, one cAMP and cGMP dependent protein kinase phosphorylation site, three N-glycosylation sites, four protein kinase C phosphorylation sites and six casein kinase 鈪，
本文编号：2245553
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