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| Tat蛋白与Plk1相互作用位点的鉴定 | |
| 引用本文： | 杨天一,张士猛,尚增甫,刘晓丹,周平坤. Tat蛋白与Plk1相互作用位点的鉴定[J]. 军事医学科学院院刊, 2011, 35(10): 749-753 |
| 作者姓名： | 杨天一 张士猛 尚增甫 刘晓丹 周平坤 |
| 作者单位： | 军事医学科学院放射与辐射医学研究所,北京,100850 |
| 基金项目： | 国家973计划项目(2007CB914603); 国家自然科学基金资助项目(30970677) |
| 摘 要： | 目的确定Tat蛋白与Plk1的相互作用及作用位点。方法构建tat和plk1基因的不同表达载体,通过GST-pull down、免疫共沉淀检测它们表达产物的相互作用位点,激光共聚焦确定其在细胞内的定位。结果 GST-pull down和免疫共沉淀实验发现Tat蛋白能结合在Plk1 N端的激酶区域,激光共聚焦发现Tat蛋白和Plk1在细胞核内有相同的亚细胞定位。结论 Tat蛋白能结合在Plk1 N端的激酶区域引起Plk1 T210的磷酸化水平提高,从而激活Plk1的活性,导致细胞周期的紊乱。 |
| 关 键 词： | 基因产物,Tat Plk1 基因表达 显微镜检查,共聚焦 细胞周期 转染 |
Identification of the interaction site of Tat and Plk1 |
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| YANG Tian-yi,ZHANG Shi-meng,SHANG Zeng-fu,LIU Xiao-dan,ZHOU Ping-kun. Identification of the interaction site of Tat and Plk1[J]. Bulletin of the Academy of Military Medical Sciences, 2011, 35(10): 749-753 | |
| Authors: | YANG Tian-yi ZHANG Shi-meng SHANG Zeng-fu LIU Xiao-dan ZHOU Ping-kun |
| Affiliation: | YANG Tian-yi,ZHANG Shi-meng,SHANG Zeng-fu,LIU Xiao-dan,ZHOU Ping-kun*(Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China) |
| Abstract: | ObjectiveTo identify the interaction site of Tat with Plk1.MethodsDifferent expression vectors of tat and plk1 were constructed.The interaction of tat and plk1 was determined by GST-pull down and immunoprecipitation assay.The localization of Tat and Plk1 was observed by the confocal laser scanning microscopy.ResultsTat protein could interact with the amino-terminal kinase domain of Plk1 as shown in GST-pull down and immunoprecipitation assay.The co-localiation of Tat and Plk1 in the nuclei of HeLa and 293 c... |
| Keywords: | gene products Tat Plk1 gene expression microscopy confocal cell cycle transfection |
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