title: 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 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.
authors.affiliation: Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, P. R. China.
authors.forename: Zhikang
authors.initials: Z
authors.lastname: Qian
authors.forename: Baoqin
authors.initials: B
authors.lastname: Xuan
authors.forename: Jie
authors.initials: J
authors.lastname: Hong
authors.forename: Zhaojin
authors.initials: Z
authors.lastname: Hao
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.forename: Weida
authors.initials: W
authors.lastname: Huang
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics*; D058619:Maltose-Binding Proteins; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D043244:Ribonuclease III / Q000096:biosynthesis* / Q000302:isolation & purification*; D012568:Schizosaccharomyces / Q000201:enzymology*
chemical_list: D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D012330:RNA, Double-Stranded; D011993:Recombinant Fusion Proteins; D043244:Ribonuclease III