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| GST pull-down验证流感病毒PB1-F2蛋白与热休克蛋白40的相互作用 | |
| 引用本文： | 侯佩莉,关振宏,张茂林,李影,段铭.GST pull-down验证流感病毒PB1-F2蛋白与热休克蛋白40的相互作用[J].生物技术通报,2011(1). |
| 作者姓名： | 侯佩莉 关振宏 张茂林 李影 段铭 |
| 作者单位： | 1. 吉林农业大学动物科学技术学院,长春,130118;吉林大学人兽共患病研究所,人兽共患病研究教育部重点实验室,长春,130062 2. 吉林大学人兽共患病研究所,人兽共患病研究教育部重点实验室,长春,130062 3. 吉林农业大学动物科学技术学院,长春,130118 |
| 基金项目： | 国家自然科学基金项目(30800824); 吉林大学农学部人才引进启动基金项目(4305050102B9) |
| 摘 要： | 为体外验证流感病毒PB1-F2与热休克蛋白Hsp40相互作用,通过两个方向的GST pull-down试验验证PB1-F2与Hsp40的相互作用。构建GST-多肽融合蛋白原核表达载体pGEX-6P-1-PB1-F2和pGEX-6P-1-Hsp40,并在大肠杆菌(E.co-li)BL21中诱导表达;构建真核表达载体pLEGFP-Hsp40及pCAGGS-PB1-F2,并分别转染293T细胞使其表达Hsp40及PB1-F2融合蛋白,然后进行GST pull-down试验验证二者的相互作用。成功地构建了两种蛋白的各种表达载体,经表达、纯化获得了可溶性的GST-多肽融合蛋白,GST pull-down试验正反两方向都证实了PB1-F2与Hsp40的相互作用,初步证实了流感病毒PB1-F2在体外能与Hsp40发生相互作用。 |
| 关 键 词： | GST pull-down 流感病毒PB1-F2蛋白 热休克蛋白40 蛋白质相互作用 |
Identification of Interaction between Influenza Virus PB1-F2 Protein and Heat Shock Protein 40 by GST Pull-down Assay |
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| Hou Peili,Guan Zhenhong,Zhang Maolin,Li Ying,Duan Ming.Identification of Interaction between Influenza Virus PB1-F2 Protein and Heat Shock Protein 40 by GST Pull-down Assay[J].Biotechnology Bulletin,2011(1). | |
| Authors: | Hou Peili Guan Zhenhong Zhang Maolin Li Ying Duan Ming |
| Institution: | Hou Peili1,2 Guan Zhenhong2 Zhang Maolin2 Li Ying1 Duan Ming2 (1College of Animal Science and Technology,Jilin Agricultral University,Changchun 130118,2Key Laboratory ofZoonosis Ministry of Education,Institute of Zoonosis,Jilin University,Changchun 130062) |
| Abstract: | To identify the interaction between influenza virus PB1-F2 protein and heat shock protein Hsp40 by GST pull-down assay in vitro,the interaction of PB1-F2 and Hsp40 was confirmed by double-direction GST pull-down assays.The prokaryotic expression vectors of GST polypeptide fusion protein pGEX-6P-1-PB1-F2 and pGEX-6P-1-Hsp40 were constructed and transformed to E.coli BL21,and then induced to express the fusion proteins.The eukaryotic vector pLEGFP-Hsp40 and pCAGGS-PB1-F2 were constructed and transfected into ... |
| Keywords: | GST pull-down Influenza virus PB1-F2 protein Hsp40 Protein-protein interaction |
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