小鼠Bicc1基因RNAi序列的筛选与鉴定
文献类型：期刊论文
| 作者 | 周亮1,2; 杨君兴[*]1 |
| 刊名 | 动物学研究
 |
| 出版日期 | 2010 |
| 卷号 | 31期号:1页码:84-88 |
| 关键词 | 小鼠Bicc1基因 RNA干扰 pRS-Hush pEGFP-C3-Bicc1 转染 |
| 其他题名 | Screening and Identification of Mouse Bicc1 RNAi |
| 通讯作者 | yangjx@mail.kiz.ac.cn |
| 合作状况 | 其它 |
| 中文摘要 | 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3个干涉序列片段后克隆到pRS-HushshRNA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Westernblotting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。 |
| 英文摘要 | 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 could knock-down the Bicel expression levels in HEK-293cells significantly were confirmed by cell immunofluorescent staining, semi-quantitative PCR and Western blotting. The results demonstrate that we have successfully obtained two efficent Bicc1 RNAi sequences, which lays a foundation for further studying on the construction of Bicc1 knock-down stable cell lines and biological function of mouse Bicc 1 product. |
| 收录类别 | 其他 |
| 语种 | 中文 |
| 公开日期 | 2010-08-19 |
| 源URL | [http://159.226.149.42:8088/handle/152453/1716]  |
| 专题 | 昆明动物研究所_系统进化与生物地理学 |
| 作者单位 | 1.中国科学院昆明动物研究所;云南昆明650223 2.中国科学院研究生院;北京100049 |
| 推荐引用方式 GB/T 7714 | 周亮,杨君兴[*]. 小鼠Bicc1基因RNAi序列的筛选与鉴定[J]. 动物学研究,2010,31(1):84-88. |
| APA | 周亮,&杨君兴[*].(2010).小鼠Bicc1基因RNAi序列的筛选与鉴定.动物学研究,31(1),84-88. |
| MLA | 周亮,et al."小鼠Bicc1基因RNAi序列的筛选与鉴定".动物学研究 31.1(2010):84-88. |
入库方式： OAI收割
来源：昆明动物研究所
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