A modified procedure for fast purification of T7 RNA polymerase.
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.
Li
Y
Y
State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, Academia Sinica, Shanghai, 200031, China.
Wang
E
E
Wang
Y
Y
D002852:Chromatography, Ion Exchange; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification; D014764:Viral Proteins
D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases