猪ANK1新的可变剪接体功能预测、组织表达谱分析与真核表达载体构建

发布时间：2018-03-25 01:29

本文选题：猪　切入点：ANK基因　出处：《基因组学与应用生物学》2017年07期

【摘要】：为进一步了解猪ANK1基因,本研究以广西巴马猪作为实验对象,分析ANK1基因结构与功能,并利用实时荧光定量PCR方法,分析ANK1基因在广西巴马小型猪11个不同组织(心脏,肝脏,脾脏,肺脏,肾脏,大肠,小肠,胰腺,皮下脂,胃,背最长肌)中的表达水平。组织表达谱分析结果表明ANK1基因在11个组织中均有表达,mRNA表达量表现为:胰腺心脏背最长肌胃大肠肝脏肾脏小肠脾脏肺脏脂肪。ANK1基因编码的蛋白通过生物信息学分析表明,编码156个氨基酸,存在跨膜结构和跨膜信号肽,分子量为17 799.27,理论等电点6.31,α螺旋和无规则卷曲所占比例比较大。通过酶切技术成功获得p EGFP-ANK1重组质粒并转染到C2C12细胞中,于48 h拍照发现细胞发出荧光,说明ANK1基因真核表达载体构建成功。
[Abstract]:In order to further understand pig ANK1 gene, this study analyzed the structure and function of ANK1 gene in Guangxi Bama pig, and analyzed ANK1 gene in 11 tissues (heart, liver) of Guangxi Bama mini-pig by real-time fluorescence quantitative PCR. Spleen, lung, kidney, large intestine, small intestine, pancreas, subcutaneous fat, stomach, The expression of ANK1 gene in 11 tissues was as follows: pancreas, heart, longissimus, large intestine, liver, kidney, small intestine, spleen, lung, fat. ANK1 gene. A bioinformatics analysis of the encoded protein shows that, Encoding 156 amino acids with transmembrane structure and transmembrane signal peptide, the molecular weight is 17 799.27, the theoretical isoelectric point is 6.31, the proportion of 伪 helix and irregular curl is relatively large. The recombinant plasmid of p EGFP-ANK1 was successfully obtained by enzyme digestion and transfected into C2C12 cells. The fluorescence of the cells was observed at 48 h, indicating that the eukaryotic expression vector of ANK1 gene was successfully constructed.
【作者单位】：广西大学动物科学技术学院;
【基金】：国家现代农业技术体系广西生猪创新团队项目(nycytxgxcxtd-03-15)资助
【分类号】：Q78;S828

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