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| 小鼠Uncv基因克隆及真核表达 | |
| 引用本文： | 徐嫄,刘冰,李文龙,胡仲明,曾林,何聪芬. 小鼠Uncv基因克隆及真核表达[J]. 中国实验动物学报, 2012, 20(1): 81-83 |
| 作者姓名： | 徐嫄 刘冰 李文龙 胡仲明 曾林 何聪芬 |
| 作者单位： | 1. 北京工商大学,北京 100048;军事医学科学院实验动物中心,北京100071 2. 军事医学科学院实验动物中心,北京,100071 3. 北京工商大学,北京,100048 |
| 基金项目： | 国家自然科学基金重点项目（编号：31030058）. |
| 摘 要： | 目的 克隆小鼠的Uncv基因并在真核细胞表达.方法 采用RT-PCR方法扩增小鼠皮肤组织中Uncv基因编码区,以真核表达质粒pcDNA 3.1-Flag为载体,构建Uncv真核表达质粒,将重组载体转染Hela细胞并用Western blot法检测基因表达.结果 构建Uncv基因真核表达载体pcDNA 3.1-Flag/Unev,重组质粒在Hela细胞中有效表达约95×103的融合蛋白.结论 成功构建真核表达载体pcDNA 3.1-Flag/Uncv,并且在真核细胞中有效表达,为研究Uncv基因生物学功能奠定基础. |
| 关 键 词： | Uncv 克隆 真核表达 毛囊 |
Cloning and eukaryotic expression of mouse Uncv gene |
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| XU Yuan , LIU Bing , LI Wen-long , HU Zhong-ming , ZENG Lin , HE Cong-fen. Cloning and eukaryotic expression of mouse Uncv gene[J]. Acta Laboratorium Animalis Scientia Sinica, 2012, 20(1): 81-83 | |
| Authors: | XU Yuan LIU Bing LI Wen-long HU Zhong-ming ZENG Lin HE Cong-fen |
| Affiliation: | 1. Beijing Technology and Business University, Beijing 100048, China; 2. Laboratory Animal Center of the Academy of the Military Medical Sciences, Beijing 100071 ) |
| Abstract: | Objective To clone the full length cDNA of Uncv gene in mice and to express the gene in eukaryotic cells. Methods RT-PCR assay was applied to clone the full length coding region of the Uncv gene and constructed its expression plasmid pcDNA 3. 1-Flag/Uncv. The recombinant plasmid was transfected into HeLa cells and the fusion protein was identified by Western blot analysis. Results The complete coding sequence was obtained and cloned into the pcDNA 3.1-Flag vector. The recombinant pcDNA 3.1-Flag/Uncv plasmid was transiently expressed in HeLa cells. HeLa cell clones expressing fusion protein with molecular weight of about 95 x 103 were obtained. Conclusions A recombinant eukaryotic expression plasmid of Uncv has been successfully constructed and expressed in HeLa cells. It may provide a foundation for further biological studies of Uncv gene. |
| Keywords: | Uncv clone eukaryotic expression hair follicle |
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