T7 Rna Polymerase Promoter

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T7 RNA polymerase - Wikipedia

(2 days ago) In biotechnology applications, T7 RNA polymerase is commonly used to transcribe DNA that has been cloned into vectors that have two (different) phage promoters (e.g., T7 and T3, or T7 and SP6) in opposite orientation. RNA can be selectively synthesized from either strand of the insert DNA with the different polymerases.

https://en.wikipedia.org/wiki/T7_RNA_polymerase

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What is the promoter sequence of T7 RNA Polymerase? | NEB

(2 days ago) T7 RNA polymerase starts transcription at the underlined G in the promoter sequence. The polymerase then transcribes using the opposite strand as a template from 5’->3’. The first base in the transcript will be a G.

https://www.neb.com/faqs/2015/01/30/what-is-the-promoter-sequence-of-t7-rna-polymerase

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T7 RNA Polymerase (20 U/µL)

(3 days ago) Thermo Scientific Bacteriophage T7 RNA polymerase is a DNA-dependent RNA polymerase with strict specificity for its respective double-stranded promoters. It catalyzes the 5'3' synthesis of RNA on either single-stranded DNA or double-stranded DNA downstream from it promoter.Highlights Incorporates mo

https://www.thermofisher.com/order/catalog/product/EP0111

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T7 RNA polymerase: promoter structure and polymerase ...

(11 months ago) A specific, promoter-independent activity of T7 RNA polymerase suggests a general model for DNA/RNA editing in single subunit RNA Polymerases. Scientific Reports 2018 , 8 (1) DOI: 10.1038/s41598-018-32231-6.

https://pubs.acs.org/doi/abs/10.1021/bi00570a023

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Promoter specificity determinants of T7 RNA polymerase | PNAS

(1 months ago) The high specificity of T7 RNA polymerase (RNAP) for its promoter sequence is mediated, in part, by a specificity loop (residues 742–773) that projects into the DNA binding cleft (1). Previous work demonstrated a role for the amino acid residue at position 748 (N748) in this loop in discrimination of the base pairs (bp) at positions −10 and −11 (2).

https://www.pnas.org/content/95/2/515

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Expression using the T7 RNA polymerase/promoter system

(2 months ago) T7 RNA polymerase is also highly selective for initiation at its own promoter sequences and is resistant to antibiotics such as rifampicin that inhibit E. coli RNA polymerase. Consequently, the addition of rifampicin to cells that are producing T7 RNA polymerase results in the exclusive expression of genes under the control of a T7 RNA ...

https://pubmed.ncbi.nlm.nih.gov/18265127/

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A mutation in T7 RNA polymerase that facilitates promoter ...

(7 days ago) Like multisubunit RNA polymerases (RNAPs), T7 RNAP frequently releases its transcript over the initial 8-12 transcribed nucleotides, when it still contacts the promoter. This abortive cycling, which is most prominent with initial sequences that deviate from those of T7 late genes, eventually comprom …

https://pubmed.ncbi.nlm.nih.gov/15831591/

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A bacteriophage T7 RNA polymerase/promoter system for ...

(3 years ago) The RNA polymerase gene of bacteriophage T7 has been cloned into the plasmid pBR322 under the inducible control of the lambda PL promoter. After induction, T7 RNA polymerase constitutes 20% of the soluble protein of Escherichia coli, a 200-fold increase over levels found in T7-infected cells.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC397196/

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Expression Using the T7 RNA Polymerase/Promoter System

(1 months ago) T7 RNA polymerase is a very active enzyme: it synthesizes RNA at a rate several times that of E. coli RNA polymerase and it terminates transcription less frequently; in fact, its transcription can ...

https://www.researchgate.net/publication/5587260_Expression_Using_the_T7_RNA_PolymerasePromoter_System

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Addgene: pCAG-T7pol

(4 days ago) T7 RNA polymerase Species. Bacteriophage T7 Insert Size (bp) 2652 Promoter CAG Cloning Information Cloning method Unknown 5′ sequencing primer CTCTAGAGCCTCTGCTAACc 3′ sequencing primer CAGAAGTCAGATGCTCAAG (Common Sequencing Primers) Resource Information. Terms and Licenses ...

https://www.addgene.org/59926/

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Building an Inducible T7 RNA Polymerase/T7 Promoter ...

(6 days ago) To develop tightly regulated orthogonal gene expression circuits in the photoautotrophic cyanobacterium Synechocystis sp. PCC6803 (Syn6803), we designed a circuit in which a native inducible promoter drives the expression of phage T7 RNA polymerase (T7RNAP). T7RNAP, in turn, specifically recognizes the T7 promoter that is designed to drive GFP expression. In Syn6803, this T7RNAP/T7promoter-GFP ...

https://pubs.acs.org/doi/10.1021/acssynbio.8b00515

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A mutation in T7 RNA polymerase that facilitates promoter ...

(1 months ago) Like multisubunit RNA polymerases (RNAPs), T7 RNAP frequently releases its transcript over the initial 8-12 transcribed nucleotides, when it still contacts the promoter. This abortive cycling, which is most prominent with initial sequences that deviate from those of T7 late genes, eventually compromises productive transcription. Starting from an in vivo situation where transcription of a ...

https://www.pnas.org/content/102/17/5958

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Ambion™ T7 RNA Polymerase, cloned, 200 U/µL

(3 days ago) This cloned high-purity Ambion T7 RNA Polymerase has a high specificity for its promoter. The enzyme will transcribe large amounts of RNA from DNA sequences downstream of the promoter and show no cross talk between promoters. One tube containing 30,000 U at a concentration of 200 U/ L is provided. I

https://www.thermofisher.com/order/catalog/product/AM2085

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Sauer:In vitro transcription with T7 RNA polymerase ...

(4 days ago) The polymerase needs something to bind to. It is a good idea to have a generic T7 promoter primer that you can use to PCR any template that has the promoter. The one I use has the sequence 5´-GAA ATT AAT ACG ACT CAC TAT A-3´ (promoter sequence in bold). This primer is also useful for sequencing plasmids that have the T7 RNAP promoter.

https://openwetware.org/wiki/Sauer:In_vitro_transcription_with_T7_RNA_polymerase

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T7 RNA Polymerase | NEB

(2 days ago) Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology.

https://www.neb.com/products/m0251-t7-rna-polymerase

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T7 RNA Polymerase - an overview | ScienceDirect Topics

(4 days ago) In T7 RNA polymerase, this conserved core is accessorized with modules that include the N-terminal domain, which is involved in the RNA polymerase-specific functions of promoter recognition and opening, RNA binding and displacement, and transcription bubble resolution and transcription termination.

https://www.sciencedirect.com/topics/neuroscience/t7-rna-polymerase

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Transcription of RNA templates by T7 RNA polymerase ...

(3 days ago) T7 RNA polymerase is processive on an RNA template. As previously observed with T3 RNA polymerase ( 19), T7 RNA polymerase is found to be processive on an RNA template following initiation at a double stranded promoter. The transition from DNA to RNA in the template strand at +18 was efficiently bypassed by the T7 RNA polymerase elongation ...

https://academic.oup.com/nar/article/26/15/3550/2360082

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T7 promoter Sequence and Map

(2 days ago) Promoter for bacteriophage T7 RNA polymerase. The map, notes, and annotations on this page and in the sequence/map file are copyrighted material.

https://www.snapgene.com/resources/plasmid-files/?set=basic_cloning_vectors&plasmid=T7_promoter

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T7 RNA Polymerase (High Concentration) | NEB

(2 days ago) Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 kD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology. Product Source

https://international.neb.com/products/m0460-t7-rna-polymerase-high-concentration

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T7 RNA Polymerase Protocol - Promega

(7 days ago) T7 RNA Polymerase is a DNA-dependent phage RNA polymerase that exhibits extremely high specificity for its cognate promoter sequence. T7 RNA Polymerase does not recognize SP6 or T3 RNA Polymerase promoter sequences as a start site for transcription. T7 RNA Polymerase is available in Promega's RiboMAX™ and Riboprobe® Systems.

https://www.promega.com/resources/protocols/product-information-sheets/n/t7-rna-polymerase-protocol/

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T7 RNA Polymerase - Promega

(5 days ago) T7 RNA Polymerase is a DNA-dependent RNA polymerase that exhibits extremely high specificity for its cognate promoter sequences. Only T7 DNA or DNA cloned downstream from a T7 promoter can serve as a template for T7 RNA Polymerase-directed RNA synthesis.

https://www.promega.com/products/cloning-and-dna-markers/molecular-biology-enzymes-and-reagents/t7-rna-polymerase/?catNum=P2075

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T7 RNA Polymerase from Escherichia coli BL 21/pAR 1219 ...

(2 days ago) Although the T7 and T3 promoter sequences differ only by 3 bp, T7 RNA polymerase only transcribes DNA cloned downstream of its promoter. Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or heat to 65 °C. Application • T7 RNA Polymerase can transcribe RNA from cloned DNA templates that are downstream from a T7 ...

https://www.sigmaaldrich.com/catalog/product/roche/rpolt7ro

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Expression Using the T7 RNA Polymerase/Promoter System ...

(5 days ago) T7 RNA polymerase is also highly selective for initiation at its own promoter sequences and is resistant to antibiotics such as rifampicin that inhibit E. coli RNA polymerase. Consequently, the addition of rifampicin to cells that are producing T7 RNA polymerase results in the exclusive expression of genes under the control of a T7 RNA ...

https://currentprotocols.onlinelibrary.wiley.com/doi/full/10.1002/0471142727.mb1602s11

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Addgene: Promoters

(3 days ago) RNA polymerase will bind to this core promoter region stably and transcription of the template strand can initiate. The TATA box is a DNA sequence (5'-TATAAA-3') within the core promoter region where general transcription factor proteins and histones can bind.

https://www.addgene.org/mol-bio-reference/promoters/

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Plasmids 101: The Promoter Region – Let's Go!

(4 days ago) T7: in vitro transcription/ general expression: Promoter from T7 bacteriophage: Constitutive, but requires T7 RNA polymerase. When used for in vitro transcription, the promoter drives either the sense OR antisense transcript depending on its orientation to your gene. T7lac: High levels of gene expression: Promoter from T7 bacteriophage plus lac ...

https://blog.addgene.org/plasmids-101-the-promoter-region

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Thermostable Variants of T7 RNA Polymerase - New England ...

(5 days ago) As would be apparent, RNA polymerase variants described herein have RNA polymerase activity and, as such, can catalyze the formation of RNA in the 5′3′ direction using a DNA template. T7 RNA polymerase is a promoter-specific polymerase that transcribes downstream of a suitable promoter (e.g., TAATACGACTCACTATAG; SEQ ID NO:2).

https://www.freepatentsonline.com/y2017/0247670.html

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The T7 Expression System - Restriction Enzyme - RR School ...

(7 days ago) This is the RNA polymerase encoded by bacteriophage T7 is different from its E. coli counterpart. Unlike the a2¡2 subunit structure of the E. coli enzyme, T7 RNA polymerase is a single-subunit enzyme that binds to distinct DNA 17 bp promoter sequences (5'-TAATACGACTCACTATA-3') found upstream of the T7 viral gene it activates.

https://www.rrnursingschool.biz/restriction-enzyme/the-t7-expression-system.html

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Expression Using the T7 RNA Polymerase/Promoter System ...

(2 days ago) The second contains the T7 RNA polymerase gene under the control of a heat‐inducible E. coli promoter. Upon heat induction, the T7 RNA polymerase is produced and initiates transcription on the expression vector, resulting in turn in the expression of the gene(s) under the control of p T7.

https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/0471142727.mb1602s11

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3′ end additions by T7 RNA polymerase are RNA self ...

(1 months ago) To test whether primer extension occurs independent of promoter DNA, we (chemically) synthesized the expected 24-base RNA and incubated it with T7 RNA polymerase, in the absence of T7 promoter DNA. The synthetic RNA has same sequence as encoded by the DNA template above (with the only difference being that the sequence from position +3 to +7 is ...

https://academic.oup.com/nar/article/46/18/9253/5096072

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Extended Upstream A-T Sequence Increases T7 Promoter ...

(5 months ago) Bacteriophage T7 RNA polymerase (RNAP) 2 and T7 promoters constitute a model system for studying the protein-DNA interactions that occur during transcription as well as to understand the basic catalytic mechanism of DNA transcription. The 99-kDa single subunit enzyme of phage T7 shares many functional characteristics of transcription catalysis with multisubunit RNAPs despite lacking proteins ...

http://www.jbc.org/content/280/49/40707.full

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Bacteriophage T7 - parts.igem.org

(3 days ago) These promoters are recognized by the T7 RNA Polymerase and are constitutive meaning that their activity is dependent on the availability of RNA polymerase, but is not affected by any transcription factors. T7 promoters are known to have high activity levels and are also highly orthogonal to bacterial promoters since the respective RNA Polymerases are unrelated.

http://parts.igem.org/Bacteriophage_T7

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In vitro transcription with T7 RNA polymerase - OpenWetWare

(3 days ago) Synthesis can be scaled up for microinjection, viral RNA infection studies, in vitro translation, and binding experiments. The Sauer lab has an excellent, detailed protocol: Sauer:In vitro transcription with T7 RNA polymerase. For a detailed description of PCR-based attachment of T7 promoters see Making RNA probes with T7 transcription. See also

https://openwetware.org/wiki/In_vitro_transcription_with_T7_RNA_polymerase

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Promoter Clearance by T7 RNA Polymerase

(1 months ago) Abstract. Footprinting, fluorescence, and x-ray structural information from the initial, promoter-bound complex of T7 RNA polymerase describes the very beginning of the initiation of transcription, whereas recent fluorescence and biochemical studies paint a preliminary picture of an elongation complex.

http://www.jbc.org/lookup/doi/10.1074/jbc.M108856200

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lacUV5 - Wikipedia

(6 days ago) Modern lacUV5 is seen in the BL21(DE3) strain, which carries both a lac operon with the standard promoter and a lacUV5 operon split by the DE3 prophage (and as a result driving the T7 RNA polymerase instead). The two important mutations are underlined.

https://en.wikipedia.org/wiki/LacUV5

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RCSB PDB - 1CEZ: CRYSTAL STRUCTURE OF A T7 RNA POLYMERASE ...

(3 days ago) Although the single-polypeptide-chain RNA polymerase from bacteriophage T7 (T7RNAP), like other RNA polymerases, uses the same mechanism of polymerization as the DNA polymerases, it can also recognize a specific promoter sequence, initiate new RNA chains from a single nucleotide, abortively cycle the synthesis of short transcripts, be regulated by a transcription inhibitor, and terminate ...

https://www.rcsb.org/structure/1cez

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Regulation of promoter selection by the bacteriophage T7 ...

(3 years ago) During bacteriophage T7 infection a phage-specified RNA polymerase transcribes the late phage genes in two temporal classes (class II and class III). In this report, we show that the purified phage polymerase discriminates between the class II and class III promoters in vitro as a function of variables that alter the stability of the DNA helix.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC324390/

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Structure of a Transcribing T7 RNA Polymerase Initiation ...

(2 days ago) The structure of a T7 RNA polymerase (T7 RNAP) initiation complex captured transcribing a trinucleotide of RNA from a 17–base pair promoter DNA containing a 5-nucleotide single-strand template extension was determined at a resolution of 2.4 angstroms. Binding of the upstream duplex portion of the promoter occurs in the same manner as that in the open promoter complex, but the single-stranded ...

https://science.sciencemag.org/content/286/5448/2305.full

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T7 and SP6 RNA polymerase, which has the higher efficiency?

(3 days ago) T7 and SP6 RNA polymerase, which has the higher efficiency? Using the pGEM-T easy vector, I want to choose the better RNA polymerase ... - we sequenced our plasmid of interest and the T7 promoter ...

https://www.researchgate.net/post/T7_and_SP6_RNA_polymerase_which_has_the_higher_efficiency

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T7 RNA Polymerase - Promega

(3 days ago) SP6/T7 または T3/T7 システムの各組合わせシステムより高比活性の RNA プローブまたは転写物を合成. P1450, P1460. T7 RiboMAX™ Express Large-Scale RNA Production System. 30分以内に 5–8.5mg/ml の RNA を合成できるハイスピードシステム. P1320

https://www.promega.jp/products/cloning-and-dna-markers/molecular-biology-enzymes-and-reagents/t7-rna-polymerase/?catNum=P2075

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New England Biolabs (UK) Ltd - T7 RNA Polymerase

(16 days ago) Description. Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology.

https://www.neb.uk.com/product/m0251

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T7 Rna Polymerase | New England Biolabs | Bioz

(1 months ago) t7 rna polymerase promoter is required for transcription of the RNA of interest. The Illumina sequencing primer is used for sequencing the DNA template of the RNA of interest and serves as a docking site for the T7 RNA polymerase when it is halted. Tus protein binds to Ter site and halts the transcribing RNA polymerase.

https://www.bioz.com/result/t7%20rna%20polymerase/product/New%20England%20Biolabs

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New England Biolabs (UK) Ltd - T7 RNA Polymerase (High ...

(12 days ago) Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 kD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology. Product Source

https://www.neb.uk.com/products/neb-catalogue/rna/t7-rna-polymerase-(high-concentration)

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A novel analytical principle using AP site-mediated T7 RNA ...

(6 days ago) T7 RNA polymerase is highly promoter-specific and only transcribes DNA downstream of the T7 promoter. We have found that modifying the T7 promoter sequence with an AP site can regulate T7 RNA polymerase transcription ability according to different modification sites. In the binding region of the promoter, AP sites greatly inhibit transcription.

https://pubs.rsc.org/en/content/articlelanding/2020/an/d0an00509f#!

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T3 and T7 RNA polymerase, Bacteriophage T7 RNA Polymerase ...

(3 days ago) Bacteriophage T3 and T7 RNA polymerases are DNA-dependent RNA polymerases with high sequence specificity for T3 or T7 promoters. Both T3 and T7 RNA polymerases synthesize RNA 5' to 3' and incorporate 35 S, 32 P, and 33 P ribonucleotides. These enzymes are isolated from an overproducing recombinant E. coli clone.

https://www.agilent.com/en/product/polymerase-chain-reaction-(pcr)/pcr-enzymes-reagents/rna-polymerases-for-pcr/t3-t7-polymerases-785935

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