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PMID:15907173

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Citation

Qian, Z, Xuan, B, Hong, J, Hao, Z, Wang, L and Huang, W (2005) Expression and purification of the carboxyl terminus domain of Schizosaccharomyces pombe dicer in Escherichia coli. Protein Pept. Lett. 12:311-4

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.

Links

PubMed

Keywords

Carrier Proteins/chemistry; Chromatography, Affinity; Escherichia coli/genetics; Maltose-Binding Proteins; RNA Interference; RNA, Double-Stranded/metabolism; Recombinant Fusion Proteins/biosynthesis; Ribonuclease III/biosynthesis; Ribonuclease III/isolation & purification; Schizosaccharomyces/enzymology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

SCHPO:DCR1

involved_in

GO:0006396: RNA processing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

SCHPO:DCR1

involved_in

GO:0030422: production of siRNA involved in RNA interference

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

SCHPO:DCR1

enables

GO:0004540: ribonuclease activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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