| 应用酵母双杂交系统筛选与NAP1相互作用的蛋白质 |
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| 引用本文： | 王烨,李峰,刘求真,谢卫兵,姚开泰. 应用酵母双杂交系统筛选与NAP1相互作用的蛋白质[J]. 湖南文理学院学报(自然科学版), 2008, 20(2): 52-59 |
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| 作者姓名： | 王烨 李峰 刘求真 谢卫兵 姚开泰 |
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| 作者单位： | 湖南文理学院,生命科学学院,湖南,常德,415000;湖南文理学院,生命科学学院,湖南,常德,415000;南方医科大学,肿瘤研究所,广东,广州,510515;南方医科大学,肿瘤研究所,广东,广州,510515 |
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| 基金项目： | 广东省自然科学基金 , 湖南省教育厅科研项目 , 湖南文理学院科技基金重点课题 |
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| 摘 要： | 用NAP1作为"诱饵"蛋白,通过酵母双杂交系统筛选人淋巴细胞cDNA文库,鉴定阳性克隆;再利用酵母双杂交和免疫共沉淀验证相互作用. 结果表明,通过酵母双杂交系统筛选人淋巴细胞cDNA文库,阳性克隆的鉴定,发现了与NAP1的相互作用蛋白质―蛋白酶体α亚基3(PSMA3). 免疫共沉淀(Co-IP)结果证实外源表达的NAP1蛋白和PSMA3蛋白在293T细胞中有特异性的相互作用. NAP1作为FDC分泌的一种多肽,与PSMA3有特异性的相互作用,这种相互作用如何实现对蛋白酶体降解靶蛋白的活性调节,其作用机理有待深入研究.
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| 关 键 词： | 酵母双杂交系统 免疫共沉淀 NAP1 相互作用蛋白 筛选 |
Identification of NAP1 protein by yeast two-hybrid system |
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| WANG Ye,LI Feng,,LIU Qiu-zhen,XIE Wei-bing,YAO Kai-Tai. Identification of NAP1 protein by yeast two-hybrid system[J]. Journal of Hunan University of Arts and Science:Natural Science Edition, 2008, 20(2): 52-59 |
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| Authors: | WANG Ye LI Feng LIU Qiu-zhen XIE Wei-bing YAO Kai-Tai |
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| Affiliation: | WANG Ye1,LI Feng1,2,LIU Qiu-zhen2,XIE Wei-bing2,YAO Kai-Tai2 (1. Life Science College,Hunan University of Arts , Science,Changde,Hunan,415000,2. Cancer Research Institute,Southern Medical University,Guangzhou,Guangdong,510515) |
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| Abstract: | Yeast two-hybrid method was used to screen human lymphocyte cDNA library with the bait plasmid that was constructed containing the whole ORF sequence of gene NAP1. Briefly, cDNA fragments encoding NAP1 were amplified by PCR and subsequently cloned into T-vector. After verified by sequencing, they were subcloned into the bait vector pGBKT7 of yeast two-hybrid system. These recombinant plasmids were transferred into Y190. Protein Interaction was confirmed by the method of Co-immunoprecipitation. Briefly, vect... |
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| Keywords: | yeast two-hybrid co-immunoprecipitation NAP1 protein-protein interaction screening |
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