| 徐淮山羊LPL基因克隆及其表达产物亚细胞定位的研究 |
| |
| 引用本文： | 秦玉蓉,许峰,田智泉,高波,张振韬,阴彦辉,孙敏,李碧春.徐淮山羊LPL基因克隆及其表达产物亚细胞定位的研究[J].畜牧兽医学报,2011,42(6). |
| |
| 作者姓名： | 秦玉蓉 许峰 田智泉 高波 张振韬 阴彦辉 孙敏 李碧春 |
| |
| 作者单位： | 扬州大学动物科学与技术学院,扬州,225009 |
| |
| 基金项目： | 转基因生物新品种培育重大专项 |
| |
| 摘 要： | 旨在克隆徐淮山羊脂蛋白酯酶(LPL)基因的cDNA,并通过EGFP融合蛋白实现该基因亚细胞水平的表达定位.本研究通过RT-PCR方法克隆徐淮山羊LPL基因cDNA,并初步进行生物信息学分析,构建含有增强型绿色荧光蛋白(EGFP)报告基因的融合表达载体pEGFP-C1-LPL.聚乙烯亚胺(PEI)介导pEGFP-C1-LPL转染NIH-3T3细胞,48 h后在荧光倒置显微镜下观察,并用RT-PCR方法检测LPL mRNA在细胞内的表达.结果,成功克隆出山羊LPL基因1 530 bp长的cDNA,完整阅读框大小为1437 bp,共编码478个氨基酸,GenBank登录号为GU082383.信号肽区域预测结果表明LPL蛋白含有一小段信号肽序列,其可能性为100%.信号肽酶切割位置在第23和24个氨基酸之间,其可能性达到65.9%.成功构建融合表达载体pEGFP-C1-LPL,并且RT-PCR显示转染后LPL在mRNA水平上表达明显.EGFP-LPL融合蛋白定位在NIT-3T3细胞外和细胞膜部分.结果表明,首次成功克隆出徐淮山羊LPL基因cDNA;重组质粒pEGFP-C1-LPL构建成功,并在NIH-3T3细胞中明显表达.
|
| 关 键 词： | 脂蛋白酯酶基因 真核表达 NIH-3T3细胞 绿色荧光蛋白 |
Study of Lipoprotein Lipase (LPL) Gene Cloning of Xuhuai Goat and Sub-cellular Localization of Its Expression Product |
| |
| QIN Yu-rong,XU Feng,TIAN Zhi-quan,GAO Bo,ZHANG Zhen-tao,YIN Yan-hui,SUN Min,LI Bi-chun.Study of Lipoprotein Lipase (LPL) Gene Cloning of Xuhuai Goat and Sub-cellular Localization of Its Expression Product[J].Acta Veterinaria et Zootechnica Sinica,2011,42(6). |
| |
| Authors: | QIN Yu-rong XU Feng TIAN Zhi-quan GAO Bo ZHANG Zhen-tao YIN Yan-hui SUN Min LI Bi-chun |
| |
| Institution: | QIN Yu-rong,XU Feng,TIAN Zhi-quan,GAO Bo,ZHANG Zhen-tao,YIN Yan-hui,SUN Min,LI Bi-chun(College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China) |
| |
| Abstract: | The aims of the study were to clone cDNA of lipoprotein lipase(LPL) gene of Xuhuai goat,and analyze the sub-cellular localization of the expression product through EGFP fusion protein.cDNA of LPL gene of Xuhuai goat was cloned by RT-PCR.Preliminary analysis of bioinformatics was implemented.Fusion expression vector named pEGFP-C1-LPL containing the enhanced green fluorescent protein(EGFP) was constructed.pEGFP-C1-LPL were transfected into NIH-3T3 cells through polyethylene imine(PEI) and then observed under... |
| |
| Keywords: | Lipoprotein lipase(LPL) gene eukaryotic expression NIH-3T3 cells GFP |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
