| Zif268基因突变体的克隆与真核表达载体的构建 |
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| 引用本文： | 张书祥,赵志虎.Zif268基因突变体的克隆与真核表达载体的构建[J].生物学杂志,2003,20(2):17-18,22. |
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| 作者姓名： | 张书祥 赵志虎 |
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| 作者单位： | 1. 安徽大学生命科学学院,合肥,230039
2. 军事医学科学院生物工程研究所,北京,100850 |
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| 基金项目： | 国家自然基金项目 39970 16 2 |
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| 摘 要： | 以小鼠转录因子Zif268的三锌指DNA结合区为模板，利用重叠(Overlap)PCR技术，获得Zif268关键氨基酸位点同时突变的三锌指突变体ZF123、2ZF123。以ZFl23、2ZF123为模板，PCR扩增获得TAT—ZF123，TAT—2ZF123序列。构建表达质拉PET—28—a^ —TAT—ZF123，pET—28—a^ —TAT—2ZF123。为利用HIVTAT蛋白的跨膜功能，实现ZF123、2ZF123在哺乳动物细胞中的表达打下基础。
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| 关 键 词： | 锌指蛋白 重叠PCR TAT蛋白 Zif268基因突变体 基因克隆 真核表达载体 |
| 文章编号： | 1008-9632(2003)02-0017-02 |
Cloning and construction of mutants of Zif268 DNA binding domain gene into an eukaryotic expressing vector |
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| ZHANG Shu_xiang ,ZHAO Zhi_hu.Cloning and construction of mutants of Zif268 DNA binding domain gene into an eukaryotic expressing vector[J].Journal of Biology,2003,20(2):17-18,22. |
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| Authors: | ZHANG Shu_xiang ZHAO Zhi_hu |
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| Affiliation: | ZHANG Shu_xiang 1,ZHAO Zhi_hu 2 |
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| Abstract: | Two three_finger mutants ZF123 and 2ZF123,were constructed by overlap RCRs whose templatts are the DNA binding domain of Zif268.The seguences of TAT_ZF123 and TAT_2ZF123 were obtained by other PCRs whose templates are ZF123 and 2ZF123.As a result two expressed vectors were obtained.ET_28_a +_TAT_2ZF123,whichwas benefit for the two genes.(ZF123 and 2ZF123) expressed in mammalian cell. |
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| Keywords: | Zinc_finger protein over_lap PCR TAT fusion proteins |
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