| 小鼠Biccl基因RNAi序列的筛选与鉴定 |
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| 引用本文： | 周亮,杨君兴.小鼠Biccl基因RNAi序列的筛选与鉴定[J].动物学研究,2010,31(1):84-88. |
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| 作者姓名： | 周亮 杨君兴 |
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| 作者单位： | 1. 中国科学院昆明动物研究所,云南,昆明,650223;中国科学院研究生院,北京,100049
2. 中国科学院昆明动物研究所,云南,昆明,650223 |
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| 摘 要： | 通过网上提供的生物信息学分析软件进行搜索和比对，初步筛选到3个较好的针对小鼠双尾-C（Bicc1）基因的RNA干扰（RNAi）序列。合成这三个干涉序列片段后克隆到pRS-Hush shRNA载体中。构建Bicc1基因的真核表达载体pEGFP-C3-Bicc1，将绿色荧光蛋白(GFP)标签标记在Bicc1蛋白上。利用细胞转染技术将pEGFP-C3-Bicc1与3个干涉序列载体共转染至体外培养的HEK-293细胞中，最后通过细胞荧光强度，半定量PCR和Western blotting鉴定出其中两个序列（pRS-Hush-RNAi-Bicc1-N/-C）能明显降低Bicc1蛋白在HEK-293细胞中的表达水平，为下一步建立起低表达Bicc1的稳定细胞株和研究小鼠Bicc1的功能提供了良好的材料。
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| 关 键 词： | 小鼠Bicc1基因 RNA干扰 pRS-Hush pEGFP-C3-Bicc1 转染 |
| 收稿时间： | 2009-6-10 |
Screening and Identification of Mouse Biccl RNAi |
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| ZHOU Liang,YANG Jun-Xing.Screening and Identification of Mouse Biccl RNAi[J].Zoological Research,2010,31(1):84-88. |
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| Authors: | ZHOU Liang YANG Jun-Xing |
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| Institution: | 1.Kunming Institute of Zoology;the Chinese Academy of Sciences;Kunming 650223;China;2.Graduate School of the Chinese Academy of Sciences;Beijing 100049;China |
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| Abstract: | Based on the online bioinformatics analysis and alignment results,three RNAi sequences target to the Mus musculus Bicc1 gene were obtained.The three interference fragments were synthesized and cloned into pRS-Hush shRNA Vector.The Bicc1 eukaryotic expression vector pEGFP-C3-Bicc1 was constructed,tagging the GFP to the N-terminal of the Bicc1 protein.The pEGFP-C3-Bicc1 and three pRS-Hush-RNAi were co-transfected into the cultured HEK-293 cells line,respectively.The two RNAi (pRS-Hush-RNAi-Bicc1-N/-C) that co... |
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| Keywords: | Mouse Bicc1 gene RNAi pRS-Hush pEGFP-C3-Bicc1 Transfect |
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