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

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

Source: Protein and Peptide Letters, Volume 12, Number 4, May 2005 , pp. 311-314(4)

Publisher: Bentham Science Publishers

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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.

Keywords: schizosaccharomyces pombe; dicer; mbp; rnai

Document Type: Review article

DOI: http://dx.doi.org/10.2174/0929866053765590

Affiliations: 1: Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China.

Publication date: 2005-05-01

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Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.