Expression and Purification of the Carboxyl Terminus Domain of Schizosaccharomyces pombe Dicer in Escherichia coli

Zhikang Qian; Baoqin Xuan; Jie Hong; Zhaojin Hao; Lu Wang; Weida Huang

doi:10.2174/0929866053765590

ISSN: 0929-8665
E-ISSN: 1875-5305

Expression and Purification of the Carboxyl Terminus Domain of Schizosaccharomyces pombe Dicer in Escherichia coli
Authors: Zhikang Qian¹, Baoqin Xuan², Jie Hong³, Zhaojin Hao⁴, Lu Wang⁵ and Weida Huang⁶
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¹ Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China. ² Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China. ³ Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China. ⁴ Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China. ⁵ Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China. ⁶ Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China.
Source: Protein and Peptide Letters, Volume 12, Issue 4, May 2005, p. 311 - 314
DOI: https://doi.org/10.2174/0929866053765590
- Available online: 01 May 2005

Abstract

The carboxyl terminus domain of Schizosaccharomyces pombe dicer (yDicerC) was expressed in Escherichia coli as an MBP-fusion protein (MBP-yDicerC). When the E. coli strain was cultured and induced at 25°C, the MBPyDicerC was partly expressed in the soluble fraction. It was then purified by two step affinity chromatography with amylose resin and Ni-NTA His.Bind® resin. The purified MBP-yDicerC showed double-strand RNA digestion activity. siRNA-like products about 22-nt in length were generated.

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2005-05-01

2025-12-16

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Article Type: Review Article

Keyword(s): dicer; mbp; rnai; schizosaccharomyces pombe

Expression and Purification of the Carboxyl Terminus Domain of Schizosaccharomyces pombe Dicer in Escherichia coli

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