| 广西巴马小型猪Oct4基因克隆及其在成纤维细胞中的表达 |
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| 引用本文： | 宁淑芳,李青洋,梁明明,许惠艳,杨小淦,陆阳清,卢晟盛,卢克焕.广西巴马小型猪Oct4基因克隆及其在成纤维细胞中的表达[J].兽医大学学报,2012(4):501-504,520. |
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| 作者姓名： | 宁淑芳 李青洋 梁明明 许惠艳 杨小淦 陆阳清 卢晟盛 卢克焕 |
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| 作者单位： | [1]广西亚热带生物资源保护利用重点试验室,广西南宁530004 [2]广西大学动物科学技术学院,广西南宁530004 |
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| 基金项目： | 国家自然科学基金资助项目（30860039）; 广西自然科学基金重点资助项目（2010GXNSFD013021） |
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| 摘 要： | 根据GenBank中公布的猪Oct4核苷酸序列设计合成1对引物,采用RT-PCR方法从巴马小型猪睾丸组织扩增Oct4基因编码全序列。然后将该基因重组于含有绿色荧光报告基因的真核表达载体pEGFP-N1中,构建成pEGFP-Oct4重组质粒,通过酶切电泳鉴定和DNA测序证明重组质粒构建成功。使用脂质体2000将pEGFP-Oct4转染巴马小型猪肾脏成纤维细胞,荧光显微镜下可观察到绿色荧光,转染细胞株中可检测到Oct4的转录。结果表明,成功构建了真核表达载体pEGFP-Oct4,并可在巴马小型猪肾脏成纤维细胞中表达,为进一步研究Oct4基因生物学功能奠定了基础。
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| 关 键 词： | 巴马小型猪 Oct4基因 真核表达 绿色荧光蛋白 |
Cloning of Guangxi bama minipig Oct4 gene and its expression in fibroblast cells |
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| Authors: | NING Shu-fang LI Qing-yang LIANG Ming-ming XU Hui-yan YANG Xiao-gan LU Yang-qing LU Sheng-sheng LU Ke-huan |
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| Institution: | 1.Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization,Nanning 530004,China;2 College of Animal Science and Technology,Guangxi University,Nanning 530004,China) |
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| Abstract: | The objectives of this study were to construct a eukaryotic expression vector of Oct4 gene from Guangxi bama minipig,and express the recombinant vector in fibroblast cells.One pair of primers was designed according to the Oct4 gene sequences of porcine published in GenBank.The complete coding sequences（CDS） of Oct4 was amplified from the testicle of a Guangxi bama minipig by RT-PCR.And then the Oct4 gene was inserted to the proper sites of vector pEGFP-N1 which included the green fluorescent protein gene.The recombinant eukaryotic expression vector pEGFP-Oct4 was confirmed through restriction emzyme digestion and DNA sequencing analysis.The plasmid was transfected into Guangxi bama minipig＇s kidney fibroblast cells with lipofectamineTM 2000,then the fluorescence was detected by fluorescent microscope,and the Oct4 mRNA was examined in cells with pEGFP-Oct4 transfected.The eukaryotic expression plasmid pEGFP-Oct4 was successfully constructed and Oct4 gene could be expressed in Bama minipig＇s kidney fibroblast cells.This test laid a foundation for further study on biological function of Oct4 gene. |
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| Keywords: | bama minipig Oct4 gene eukaryotic expression EGFP |
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