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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 31, No. 1, 2010, pp. 84−88
Bioline Code: zr10012
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 31, No. 1, 2010, pp. 84−88

 en Screening and Identification of Mouse Bicc1 RNAi
Zhou, Liang & Yang, Jun-Xing

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 could knock-down the Bicc1 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 Bicc1 product.

Keywords
Mouse Bicc1 gene; RNAi; pRS-Hush; pEGFP-C3-Bicc1; Transfect

 
© Copyright 2010 Kunming Institute of Zoology, the Chinese Academy of Sciences
Alternative site location: http://www.zoores.ac.cn/

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