R0884
T7 RNA Polymerase
recombinant, expressed in E. coli, buffered aqueous solution
Synonym(s):
RNA Polymerase T7, RNA Polymerase, T7 from E. coli HMS 174/pAR1219
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About This Item
Recombinant:
expressed in E. coli
Concentration:
10,000-50,000 U/mL
recombinant
expressed in E. coli
grade
Molecular Biology
form
buffered aqueous solution
mol wt
98.8 kDa
concentration
10,000-50,000 U/mL
UniProt accession no.
foreign activity
DNase and RNase, none detected
storage temp.
−20°C
Gene Information
bacteriophage T7 ... T7p07(1261050)
General description
T7 RNA polymerase is highly specific for the bacteriophage T7 promoter and terminator sequences. It is extensively used to prepare RNA transcripts for stuctural and metabolic studies. The RNA transcripts can be converted to probes for sensitive hybridization detection studies. T7 polymerase and dideoxynucleotides can be used to directly sequence DNA.
Analysis Note
Activity assay: 40 mM Tris-HCl, pH 7.9, 6 mM MgCl2, 4 mM spermidine, 10 mM DTT, 0.5 μM each rNTP + 10 μCi α-32P-UTP, 3-10 units of enzyme, and 1 μg of a 350 bp template are incubated for 10 min at 37°C in a total volume of 100 μl. Typical results are ≥50% incorporation of labeled nucleotide into ≥90% full-length transcript.
Other Notes
One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37°C.
T7 RNA Polymerase is supplied as a solution of 100 mM NaCl, 50 mM Tris-HCl (pH 7.9), 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, and 50% (v/v) glycerol.
Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
Regulatory Information
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Characterization of T7-specific ribonucleic acid polymerase. 1. General properties of the enzymatic reaction and the template specificity of the enzyme.
M Chamberlin et al.
The Journal of biological chemistry, 248(6), 2235-2244 (1973-03-25)
Ran Furman et al.
FEBS letters, 587(6), 614-619 (2013-02-19)
Transcription factor DksA contains a four-Cys Zn(2 +)-finger motif thought to be responsible for structural integrity and the relative disposition of its domains. Pseudomonas aeruginosa encodes an additional DksA paralog (DksA2) that is expressed selectively under Zn(2+) limitation. Although DksA2
Looking for a promoter in 3D.
Vladimir Svetlov et al.
Nature structural & molecular biology, 20(2), 141-142 (2013-02-06)