SP6 RNA Polymerase
SP6 RNA Polymerase
Brand: Takara Bio.
In stock
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SP6 RNA Polymerase
SP6 RNA Polymerase is a DNA-dependent RNA polymerase that exhibits a high specificity for bacteriophage SP6 promoter sequences. The enzyme can incorporate labeled or unlabeled nucleotide triphosphates into an RNA transcript. Large quantities of RNA can be synthesized from a DNA sequence cloned downstream from an SP6 promoter. SP6 RNA Polymerase is supplied in 10 mM potassium phosphate (pH 7.9), 150mM NaCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol.
Applications
- In vitro transcription
Source
Recombinant E. coli
Storage
–20°C
Unit definition
One unit is defined as the amount of enzyme required to incorporate 1 nmol of [3H] GMP into acid-insoluble product in 1 hour at 37°C and pH 7.5 using a linear DNA template.
Buffer
Supplied with 10X Reaction Buffer [400 mM Tris-HCl (pH 7.5), 60 mM MgCl2, 20 mM spermidine, 100 mM DTT, 0.1% BSA].
Concentration
10–50 U/µl
Butler, E. T. & Chamberlins, M. J. Bacteriophage SP6-specific RNA Polymerase I. ISOLATION AND CHARACTERIZATION OF THE ENZYME. J. Biol. Chem. 257, 5772–5778 (1982).
Green, M. R., Maniatis, T. & Melton, D. A. Human β-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei. Cell 32, 681–694 (1983).
Kotaoi, H., Ishizalci, Y., Hiraoka, N. & Obayashi, A. Nucleotide sequence and expression of the doned gene of bacteriophage SP6 RNA polymerase. Nucleic Acids Res. 15, (1987).
Melton, D. A. et al. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 12, 7035–56 (1984).
Pelletier, J. & Sonenberg, N. Insertion mutagenesis to increase secondary structure within the 5′ noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell 40, 515–526 (1985).
Sagawa, H., Ohshima, A. & Kato, I. A tightly regulated expression system in Escherichia coli with SP6 RNA polymerase. Gene 168, (1996).
Schenborn, E. T. & Mierendorf, R. C. A novel transcription property of SP6 and T7 RNA polymerases: dependence on template structure. Nucleic Acids Res. 13, 6223–36 (1985).
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