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R0884

Sigma-Aldrich

T7 RNA Polymerase

recombinant, expressed in E. coli, buffered aqueous solution

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Synonym(s):
RNA Polymerase T7, RNA Polymerase, T7 from E. coli HMS 174/pAR1219
CAS Number:
9014-24-8
Enzyme Commission number:
2.7.7.6 (BRENDA, IUBMB)
MDL number:
MFCD00132197

recombinant

expressed in E. coli

grade

for molecular biology

form

buffered aqueous solution

mol wt

98.8 kDa

concentration

10,000-50,000 U/mL

UniProt accession no.

P00573

foreign activity

DNase and RNase, none detected

storage temp.

−20°C

Gene Information

bacteriophage T7 ... T7p07(1261050)

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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.

Components

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.

Unit Definition

One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37°C.

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.

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Storage Class Code

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)
V D Axelrod et al.
Biochemistry, 24(21), 5716-5723 (1985-10-08)
RNA synthesis by T7 RNA polymerase or SP6 RNA polymerase is 100-1000 times more sensitive to the presence of the 3'-deoxyribonucleoside 5'-triphosphate chain terminators than is RNA synthesis by Escherichia coli RNA polymerase or Q beta replicase. These ribonucleotide analogues
Yuko Murayama et al.
Acta crystallographica. Section F, Structural biology and crystallization communications, 69(Pt 2), 174-177 (2013-02-07)
DNA-dependent RNA polymerase (RNAP) synthesizes RNA complementary to the template DNA. During transcript elongation, RNAP often undergoes backward translocation ('backtracking') by dissociating the 3' end of the nascent RNA transcript from the template DNA. While the backtracked state of RNAP
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)
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