Expression and Purification of the Carboxyl Terminus Domain of Schizosaccharomyces pombe Dicer in Escherichia coli
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.
Document Type: Review Article
Affiliations: Department of Biochemistry, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P. R. China.
Publication date: May 1, 2005
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