Product Details

Purity

Greater than 90% as determined by SDS-PAGE.

Target Names

Uniprot No.

P00573

Research Area

Signal Transduction

Alternative Names

(DNA-directed RNA polymerase)

Species

Escherichia phage T7 (Bacteriophage T7)

Source

E.coli

Expression Region

274-509aa

Target Protein Sequence

PPKPWTGITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANVITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLEFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYYWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPF
Note: The complete sequence may include tag sequence, target protein sequence, linker sequence and extra sequence that is translated with the protein sequence for the purpose(s) of secretion, stability, solubility, etc.
If the exact amino acid sequence of this recombinant protein is critical to your application, please explicitly request the full and complete sequence of this protein before ordering.

Mol. Weight

34.6 kDa

Protein Length

Partial

Tag Info

N-terminal 10xHis-tagged and C-terminal Myc-tagged

Form

Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

The expression region of this recombinant Escherichia phage T7 (Bacteriophage T7) T7 RNA polymerase covers amino acids 274-509. The expected molecular weight for the T7 RNA polymerase protein is calculated to be 34.6 kDa. This T7 RNA polymerase recombinant protein is manufactured in e.coli. The T7 RNA polymerase coding gene included the N-terminal 10xHis tag and C-terminal Myc tag, which simplifies the detection and purification processes of the recombinant T7 RNA polymerase protein in following stages of expression and purification.

The Escherichia phage T7 RNA polymerase (T7 RNAP) is an enzyme encoded by the T7 bacteriophage genome. It is a single-subunit RNA polymerase that recognizes and transcribes genes under the control of T7 promoter sequences. T7 RNAP is highly specific and efficient, making it a valuable tool in molecular biology research, particularly for in vitro transcription of RNA. Its use is prevalent in various applications, such as synthesizing RNA probes, producing RNA for structural studies, and generating messenger RNA (mRNA) for recombinant protein expression. T7 RNAP has become a standard component in molecular biology laboratories for its robustness and utility in diverse experimental techniques.

Target Background

Function

Highly processive DNA-dependent RNA polymerase that catalyzes the transcription of class II and class III viral genes. Recognizes a specific promoter sequence and enters first into an 'abortive phase' where very short transcripts are synthesized and released before proceeding to the processive transcription of long RNA chains. Unwinds the double-stranded DNA to expose the coding strand for templating. Participates in the initiation of viral DNA replication presumably by making primers accessible to the DNA polymerase, thus facilitating the DNA opening. Plays also a role in viral DNA packaging, probably by pausing the transcription at the right end of concatemer junction to allow packaging complex recruitment and beginning of the packaging process.

Gene References into Functions

Using correlation analysis, it was found that the promoter strength characterized by reporter gene expression was closely correlated with rupture force and the binding percentage of T7 RNA polymerase by force spectroscopy. PMID: 29187520
From the docking of the minimum energy representative structures of T7 lysozyme at different pH strengths (obtained from the free energy landscape analysis) with T7RNAP structures at same pH strengths, we saw strong interaction patterns at pH 7.9 and pH 5. PMID: 28545576
Data suggest that comparative transcriptional fidelities/mutation rates for DNA-directed RNA polymerases are as follows: human mitochondrial POLRMT [2x10(-5)]; Saccharomyces cerevisiae mitochondrial Rpo41 [6x10(-6)]; phage T7 single-subunit RNA polymerase [2x10(-6)]. PMID: 28882896
Molecular dynamics simulation study described the behavior of the two magnesium ions in RNAP activity during genetic transcription. PMID: 28205291
The control of the discrimination between dNTP and rNTP in DNA and RNA polymerases has been described. PMID: 27480935
The study provides evidence that the intrinsic infidelity of T7 RNAP in transcription brings significant phenotypic consequences, observed as frameshift mutation restoration and the production of a heterogenous population of the full-length protein. PMID: 25824942
We show here that DNA-protein cross-links constitute strong but not absolute blocks to in vitro transcription catalyzed by T7 RNAP. PMID: 22235136
Hydrogen bonds of RNA polymerase play an important role in the efficiency of transcription. PMID: 22044042
introducing a nuclear transgene, ST7, encoding a light-regulated plastid-targeted T7RNAP by cross-pollination. PMID: 15517992
T7 promoter-polymerase interaction weakening facilitates promoter release PMID: 15711016
An amino acid substitution weakens promoter binding but markedly reduces abortive cycling over a variety of initial sequences. PMID: 15831591
structure activity relationship PMID: 16301518
Active T7RNAP elongation by showing that both rapidly elongating and halted complexes are equally sensitive to pyrophosphate. PMID: 16516229
Collapse of the DNA from the downstream end of the bubble is a major contributor to the characteristic instability of the abortive T7 RNA polymerase transcription complex. PMID: 16790422
Poto-regulation of transcription reaction by azobenzene-tethered promoter was attributed to the change of binding property of RNAP to the promoter by trans-cis isomerization of azobenzene. PMID: 17150558
transition to an elongation complex by T7 RNA polymerase is a multistep process PMID: 17548349
site-specifically tethered chemical nucleases and functional characterization of directed T7 RNAP mutants to both reveal the architecture of the duplex DNA PMID: 17580086
G-rich sequences located in the transcribed strand do not affect transcription by either polymerase, but when the sequences are located in the non-transcribed strand, they partially arrest both polymerases PMID: 18292094
study reports crystal structures of T7 RNAP bound to promoter DNA containing either a 7- or an 8-nucleotide (nt) RNA transcript that illuminate intermediate states along the transition pathway PMID: 18948533

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Protein Families

Phage and mitochondrial RNA polymerase family

Database Links

KEGG: vg:1261050

Code	CSB-EP018353EEB
Abbreviation	Recombinant Escherichia phage T7 T7 RNA polymerase protein, partial
MSDS	MSDS Datasheet
Size	$388
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Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Recombinant Escherichia phage T7 T7 RNA polymerase (1), partial

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