| pEGFP-C3/Tsarg1融合蛋白在GC-1 spg细胞中的表达与定位 |
| |
| 引用本文： | 张懿,邢晓为.pEGFP-C3/Tsarg1融合蛋白在GC-1 spg细胞中的表达与定位[J].实用预防医学,2010,17(8):1479-1481. |
| |
| 作者姓名： | 张懿 邢晓为 |
| |
| 作者单位： | 1. 湖南省长沙市第八医院,湖南,长沙,410004
2. 中南大学湘雅三医院 |
| |
| 基金项目： | 国家自然科学基金资助项目，中国博士后基金 |
| |
| 摘 要： | 目的构建pEGFP-C3/Tsarg1真核表达载体并在GC-1spg细胞中表达,为研究Tsarg1在生精过程中的机制提供实验基础。方法应用RT-PCR从大鼠睾丸中扩增Tsarg1的开放阅读框(ORF),并将PCR产物插入到T载体中测序验证。随后,将Tsarg1cDNA片段亚克隆入载体pEGFP-C3,筛选阳性克隆,构建pEGFP-C3/Tsarg1真核表达载体。脂质体介导重组体pEGFP-C3/Tsarg1转染GC-1spg细胞,荧光显微镜观察融合蛋白在细胞内的表达与定位。结果成功构建了pEGFP-C3/Tsarg1真核表达质粒,EGFP-Tsarg1融合蛋白能够在GC-1spg细胞中表达,Tsarg1蛋白定位于细胞胞浆。结论 pEGFP-C3/Tsarg1真核表达载体的成功构建及带GFP标签的Tsarg1融合蛋白在GC-1spg细胞中的成功表达为进一步研究Tsarg1的生物学功能奠定了基础。
|
| 关 键 词： | Tsarg1 克隆 转染 表达 定位 |
Expression and Localization of pEGFP-C3/Tsarg1 Fusion Protein in GC-1 spg Cells |
| |
| ZHANG Yi,XING Xiao-wei.Expression and Localization of pEGFP-C3/Tsarg1 Fusion Protein in GC-1 spg Cells[J].Practical Preventive Medicine,2010,17(8):1479-1481. |
| |
| Authors: | ZHANG Yi XING Xiao-wei |
| |
| Institution: | ZHANG Yi,XING Xiao-wei(The Eighth Hospital of Changsha,Changsha 410004,Hunan,China) |
| |
| Abstract: | Objective To construct eukaryotic expression vector pEGFP-C3/Tsarg1 and to observe the subcellular localization in GC-1 spg cells.Methods The open reading frame(ORF)of Tsarg1 was amplified from rat testis by RT-PCR.The PCR products were cloned into pUCm-T vectors and sequenced.Then the cDNA fragment was subcloned into pEGFP-C3 vector and the positive clones were selected.The recombinant plasmid pEGFP-C3/Tsarg1 was transfected into the GC-1 spg cells using liposome.The expression and localization of the fusi... |
| |
| Keywords: | Tsarg1 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
