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A modified procedure for fast purification of T7 RNA polymerase
1State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, Academia Sinica, Shanghai, 200031, China.
Protein Expression and Purification
|July 27, 1999
Abstract:
The in vitro T7 transcription system allows one to synthesize biochemical amounts of RNA molecules functionally equivalent or similar to those transcripts normally existing at extremely low levels in vivo. In this study we described a modified method for efficient large-scale preparation of pure T7 RNA polymerase free of RNase activity from the recombinant Escherichia coli strain BL21/pAR1219 (4). The procedure, which used preparative column chromatography on DEAE-Sepharose CL-6B and Blue 3GA, was shown to be simple, rapid, and cost effective in comparison with other methods reported previously.