SP6 RNA Polymerase
SP6 RNA Polymerase is a DNA-dependent RNA polymerase that exhibits high specificity for the bacteriophage SP6 promoter sequence 5'-TAATCCACTGTGATAT-3'. The enzyme can incorporate labeled or unlabeled ribonucleoside triphosphates into an RNA transcript. Large quantities of RNA can be synthesized from a DNA sequence cloned downstream of the SP6 promoter due to the high processivity of SP6 RNA Polymerase. The enzyme is supplied in a buffer consisting of 10 mM potassium phosphate (pH 7.9), 150 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 50% glycerol.
- In vitro transcription
Recombinant E. coli
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.
Supplied with 10X Reaction Buffer [400 mM Tris-HCl (pH 7.5), 60 mM MgCl2, 20 mM spermidine, 100 mM DTT, 0.1% BSA].
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).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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