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 degrees C, the MBP-yDicerC was partly expressed in the soluble fraction. It was then purified by two step affinity chromatography with amylose resin and Ni-NTA His Bind(R) resin. The purified MBP-yDicerC showed double-strand RNA digestion activity. siRNA-like products about 22-nt in length were generated.
Qian Z, Xuan B, Hong J, Hao Z, Wang L, Huang W