| 多头绒泡菌微原质团瞬时表达系统的构建 |
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| 引用本文： | 刘士德,程彩霞,林子扬,张建华,李明华,周卓龙,田生礼,邢苗.多头绒泡菌微原质团瞬时表达系统的构建[J].生物工程学报,2009,25(6):854-862. |
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| 作者姓名： | 刘士德 程彩霞 林子扬 张建华 李明华 周卓龙 田生礼 邢苗 |
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| 作者单位： | 1. 深圳大学生命科学学院,深圳市微生物基因工程重点实验室,深圳,518060
2. 深圳大学光电子研究所,深圳,518060 |
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| 基金项目： | 国家自然科学基金项目(No.30470113);;广东省自然科学基金项目(No.04011314);;深圳市科技基金项目(No.200442)资助~~ |
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| 摘 要： | 真核生物多头绒泡菌的原质团是研究细胞周期的好材料。但尚无合适的表达体系可供选择。本研究用多头绒泡菌ardC actin基因启动子和终止子分别替换哺乳动物细胞表达质粒pDsRed1-N1的CMVIE和SV40 polyA片段,构建了多头绒泡菌红色荧光蛋白(RFP)表达质粒pXM1;用PardC-MCS-DsRed1-TardC替换pTB38表达盒PardC-hph-TardC,构建了多头绒泡菌RFP表达质粒pXM2。将多头绒泡菌转录延伸因子类似蛋白(PELF1)基因与质粒pXM2重组,构建了PELF1红色荧光融合蛋白(PELF1-RFP)表达质粒pXM2-pelf1。通过荧光显微镜和激光扫描共聚焦显微镜观察RFP表达发现,电转参数为4kV/cm(电场)、1A(电流)、70μs(电击时间)时,质粒pXM1和pXM2电转多头绒泡菌微原质团(≤500μm)后24～48h内,RFP荧光最显著;而PELF1-RFP则主要聚集在多头绒泡菌细胞核,说明本试验建立的表达系统可以用于研究特定蛋白在多头绒泡菌内的瞬时表达。
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| 关 键 词： | 多头绒泡菌 微原质团 瞬时表达 |
| 收稿时间： | 2009/1/14 0:00:00 |
Transient expression in microplasmodia of Physarum polycephalum |
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| Shide Liu,Caixia Cheng,Ziyang Lin,Jianhua Zhang,Minghua Li,Zhuolong Zhou,Shengli Tian and Miao Xing.Transient expression in microplasmodia of Physarum polycephalum[J].Chinese Journal of Biotechnology,2009,25(6):854-862. |
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| Authors: | Shide Liu Caixia Cheng Ziyang Lin Jianhua Zhang Minghua Li Zhuolong Zhou Shengli Tian and Miao Xing |
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| Institution: | Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Institute of Photoelectron, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China;Shenzhen Key Laboratory of Microbial and Genetic Engineering, College of Life Sciences, Shenzhen University, Shenzhen 518060, China |
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| Abstract: | The plasmodium of Physarum polycephalum is a suitable eukaryotic cell for cell cycle investigation,but there is no compatible transient expression system for the plasmodium.Using the promoter and terminator of ardC actin of Physarum polycephalum substituted the CMV IE and SV40 polyA of plasmid pDsRed1-N1,using cassette PardC-MCS-DsRed1-TardC substituted the cassette PardC-hph-TardC of plasmid pTB38,we constructed plasmids pXM1 and pXM2 for transient expression of red fluorescent protein(RFP) in Physarum pol... |
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| Keywords: | Physarum polycephalum microplasmodium transient expression |
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