Expression and Purification of the Carboxyl Terminus Domain of Schizosaccharomyces pombe Dicer in Escherichia coli
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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Document Type: Review Article
Affiliations: 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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