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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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
Danil Pupov et al.
The Biochemical journal, 452(2), 241-248 (2013-03-23)
Besides canonical double-strand DNA promoters, multisubunit RNAPs (RNA polymerases) recognize a number of specific single-strand DNA and RNA templates, resulting in synthesis of various types of RNA transcripts. The general recognition principles and the mechanisms of transcription initiation on these
Sangjin Kim et al.
Science (New York, N.Y.), 339(6121), 816-819 (2013-02-16)
Allostery is well documented for proteins but less recognized for DNA-protein interactions. Here, we report that specific binding of a protein on DNA is substantially stabilized or destabilized by another protein bound nearby. The ternary complex's free energy oscillates as
Katsuhiko S Murakami
The Journal of biological chemistry, 288(13), 9126-9134 (2013-02-08)
Escherichia coli RNA polymerase (RNAP) is the most studied bacterial RNAP and has been used as the model RNAP for screening and evaluating potential RNAP-targeting antibiotics. However, the x-ray crystal structure of E. coli RNAP has been limited to individual
David L Shis et al.
Proceedings of the National Academy of Sciences of the United States of America, 110(13), 5028-5033 (2013-03-13)
The construction of synthetic gene circuits relies on our ability to engineer regulatory architectures that are orthogonal to the host's native regulatory pathways. However, as synthetic gene circuits become larger and more complicated, we are limited by the small number
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