| 徐淮山羊PPAR基因克隆及其表达产物亚细胞定位的研究 |
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| 引用本文： | 秦玉蓉,许峰,高波,宋正海,范广习,李碧春. 徐淮山羊PPAR基因克隆及其表达产物亚细胞定位的研究[J]. 中国畜牧杂志, 2011, 47(7) |
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| 作者姓名： | 秦玉蓉 许峰 高波 宋正海 范广习 李碧春 |
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| 作者单位： | 1. 扬州大学动物科学与技术学院,江苏扬州,225009
2. 苏州市吴中区东山动物防疫站,江苏苏州,215128 |
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| 基金项目： | 转基因生物新品种培育重大专项(2008ZX08008-003、2009ZX08008-003B) |
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| 摘 要： | 研究旨在克隆徐淮山羊过氧化物酶体激活增殖受体(PPAR)基因的cDNA,并通过EGFP融合蛋白对该基因产物亚细胞水平定位。采用RT-PCR方法克隆徐淮山羊脂肪组织PPAR基因cDNA,并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-PPAR,聚乙烯亚胺(PEI)介导基因转染NIH-3T3细胞,48h后荧光倒置显微镜下观察基因表达产物的分布,RT-PCR检测mRNA在体外水平的表达。结果表明:首次成功克隆出徐淮山羊PPAR基因的全序列cDNA,大小为1428bp,GenBank登录号为GU082382;构建了融合表达载体pEGFP-C1-PPAR;RT-PCR检测mRNA表达明显;EGFP-PPAR融合蛋白定位在NIT-3T3细胞质中。体外克隆的徐淮山羊PPAR基因cDNA,在单细胞水平上可表达于细胞质中,结果为进一步研究PPAR的生物学功能奠定基础。
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| 关 键 词： | 过氧化氢酶体激活增殖受体 真核表达 NIH-3T3细胞 荧光蛋白 徐淮山羊 |
Study on PPAR Gene Cloning and Sub-cellular Localization of Its Expression Product in Xuhuai Goat |
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| QIN Yu-rong,XU Feng,GAO Bo,SONG Zheng-hai,FAN Guang-xi,LI Bi-chun. Study on PPAR Gene Cloning and Sub-cellular Localization of Its Expression Product in Xuhuai Goat[J]. Chinese Journal of Animal Science, 2011, 47(7) |
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| Authors: | QIN Yu-rong XU Feng GAO Bo SONG Zheng-hai FAN Guang-xi LI Bi-chun |
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| Affiliation: | QIN Yu-rong1,XU Feng1,GAO Bo1,SONG Zheng-hai2,FAN Guang-xi2,LI Bi-chun1(1.College of Animal Science and Technology,Yangzhou University,Jiangsu Yangzhou 225009,China,2.DongshanAnimal Epidemic Prevention Station of Wuzhong District,Jiangsu Suzhou 215128,China) |
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| Abstract: | The study aimed to clone cDNA of peroxisome proliferator-activated receptor(PPAR) gene of Xuhuai goat,and analysis sub-cellular localization of the expression product through EGFP fusion protein.cDNA of PPAR gene of Xuhuai goat was cloned and submitted to GenBank;fusion expression vector named pEGFP-C1-PPAR which containing the enhanced green fluorescent protein(EGFP) was constructed;NIH-3T3 cells were transfected pEGFP-C1-PPAR through polyethylene imine(PEI) and then observed under inverted microscope afte... |
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| Keywords: | peroxisome proliferator-activated receptor eukaryotic NIH-3T3 cells GFP Xuhuai goat |
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