| TAT蛋白质转导结构域与SV40启动子特异的三锌指结构融合蛋白的表达与纯化 |
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| 引用本文： | 赵兴卉,朱旭东,吴国清,黄培堂. TAT蛋白质转导结构域与SV40启动子特异的三锌指结构融合蛋白的表达与纯化[J]. 生物技术通讯, 2005, 16(1): 22-24 |
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| 作者姓名： | 赵兴卉 朱旭东 吴国清 黄培堂 |
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| 作者单位： | 军事医学科学院,生物工程研究所,北京,100071 |
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| 摘 要： | 利用蛋白质转导结构域(PTDs)可以将与之融合表达的蛋白质直接送入细胞中。将通过筛选噬菌体展示锌指库得到的特异作用于SV40启动子上9bp序列的三锌指结构的序列插入含有TAT蛋白的蛋白质转导结构域的表达载体pET—TAT-NLS中，构建融合蛋白的表达载体pET-TAT-NLS—clone3。融合蛋白在E．coli BL21(DE3)中得到了可溶性表达，含量约占总蛋白的18％；并通过镍亲和凝胶层析柱得到了较好的纯化融合蛋白。
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| 关 键 词： | TAT 蛋白质转导结构域 SV40启动子 三锌指结构 融合蛋白 可溶性表达 纯化 |
| 文章编号： | 1009-0002(2005)01-0022-03 |
| 修稿时间： | 2004-09-02 |
Expression and Purification of the Fusion Protein of TAT Protein Transduction Domain with a Zinc Finger that Specific to SV40 Promoter |
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| ZHAO Xing-hui,ZHU Xu-dong,WU Guo-qing,HUANG Pei-tangInstitute of Biotechnology,Academy of Military Medical Sciences,Beijing ,China. Expression and Purification of the Fusion Protein of TAT Protein Transduction Domain with a Zinc Finger that Specific to SV40 Promoter[J]. Letters in Biotechnology, 2005, 16(1): 22-24 |
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| Authors: | ZHAO Xing-hui ZHU Xu-dong WU Guo-qing HUANG Pei-tangInstitute of Biotechnology Academy of Military Medical Sciences Beijing China |
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| Affiliation: | ZHAO Xing-hui,ZHU Xu-dong,WU Guo-qing,HUANG Pei-tangInstitute of Biotechnology,Academy of Military Medical Sciences,Beijing 100071,China |
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| Abstract: | Protein transduction domains (PTDs), such as the PTD of TAT, may transduce peptides which fusion expressed with them into cells. A zinc finger peptide that specific to a 9 bp target on SV40 promoter was got by screening zinc finger phage display libraries. The sequence of this zinc finger was inserted into a prokaryotic expression vector named pET-TAT-NLS, generating the recombinant plasmid pET-TAT-NLS-clone3. The fusion protein was expressed in a soluable form, amount to about 18% of total proteins. The fusion protein was then purified by affinity chromatography. |
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| Keywords: | zinc finger SV40 promoter TAT protein transduction domains fusion protein |
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