pentr vector  (Thermo Fisher)


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    Name:
    Gateway pENTR 4 Dual Selection Vector
    Description:
    Gateway entry vectors are designed to clone DNA sequences using restriction endonucleases and ligase to create a Gateway entry clone The resulting entry clone is ready for recombination with a destination vector to create an expression clone The Gateway entry vectors Table 1 offer the following • attL1 and attL2 sites for site specific recombination of the entry clone with a Gateway destination vector to ensure cloning of the gene of interest in the correct orientation for expression• Kozak consensus sequence for efficient translation initiation in eukaryotic systems• Ribosome binding site for efficient translation initiation in prokaryotic systems pENTR 1A Dual Selection pENTR 3C Dual Selection and pENTR 11 Dual Selection vectors only • rrnB transcription termination sequences to prevent basal expression of the PCR product of interest in E coli • pUC origin for high copy replication and maintenance of the plasmid in E coli• Kanamycin resistance gene for selection in E coli• The ccdB⁄chloramphenicol fusion gene located between the two attL sites for o negative selection ando Chloramphenicol selection in E coli• Kanamycin resistance gene for selection in E coli
    Catalog Number:
    a10465
    Price:
    None
    Applications:
    Cloning|Entry Vectors & Kits|Gateway Cloning
    Category:
    DNA Vectors Clones Purified Nucleic Acids Libraries
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    Structured Review

    Thermo Fisher pentr vector
    Construction of artificial microRNAs (amiRNAs) targeting the conserved motifs of the tospoviral replicase sequence in the <t>pre‐miR159a</t> backbone. (A) Construction of single amiRNAs. MicroRNA159a (miR159a) processed from the pre‐miRNA159a backbone is presented as a hairpin. Forward primer amiR‐L‐F1 and reverse primer amiR‐L‐R, containing the complementary sequence to an individual amiRNA, were used to replace the miRNA159a and miRNA159a* sequences with an amiRNA targeting the A, B1, B2, C, D or E conserved motif of the L gene (replicase) of Watermelon silver mottle virus by first polymerase chain reaction (PCR). Bgl II‐F2 primer containing a Bgl II site and R primer containing a Sma I site were used in nested PCR. The pre‐amiRNA159a was digested with Bgl II/ Sma I, subcloned into the <t>pENTR</t> vector and placed downstream of the 35S promoter in a binary destination vector (destination cassette, DC) by the Gateway recombination system. (B) Strategy for triple amiRNA construction. The primer pair M13‐F/amiR159Xba‐R was used to amplify amiR‐LA (or amiR‐LB2). The primer pairs amiR159AvrII‐F/amiLB1XbaAscI‐R and amiR159AvrII‐F/amiLDXbaAscI‐R were used to amplify amiR‐LB1 and amiR‐LD, respectively. The primer pair amiR159AvrII‐F/M13‐R was used to amplify amiR‐LE or amiR‐LC. From the amplified products, the amiR‐LA or amiR‐LB2 fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LB1 or amiR‐LD fragment, respectively. After ligation, the double amiRNA segment was amplified by the primer pair M13‐F/amiLB1XbaAscI‐R or M13‐F/amiLDXbaAscI‐R. The fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LE or amiR‐LC fragment. The triple amiRNA segment was amplified by M13II‐F/pENTRAscI. The triple amiRNA fragment was released from the amplified product by Not I/ Asc I digestion and subcloned into the pENTR vector to generate pENTR‐amiR‐LAB1E and pENTR‐amiR‐LB2DC, and then into binary DC vector by Gateway recombination to form triple amiRNA constructs pre‐amiR‐LAB1E and pre‐amiR‐LB2DC.
    Gateway entry vectors are designed to clone DNA sequences using restriction endonucleases and ligase to create a Gateway entry clone The resulting entry clone is ready for recombination with a destination vector to create an expression clone The Gateway entry vectors Table 1 offer the following • attL1 and attL2 sites for site specific recombination of the entry clone with a Gateway destination vector to ensure cloning of the gene of interest in the correct orientation for expression• Kozak consensus sequence for efficient translation initiation in eukaryotic systems• Ribosome binding site for efficient translation initiation in prokaryotic systems pENTR 1A Dual Selection pENTR 3C Dual Selection and pENTR 11 Dual Selection vectors only • rrnB transcription termination sequences to prevent basal expression of the PCR product of interest in E coli • pUC origin for high copy replication and maintenance of the plasmid in E coli• Kanamycin resistance gene for selection in E coli• The ccdB⁄chloramphenicol fusion gene located between the two attL sites for o negative selection ando Chloramphenicol selection in E coli• Kanamycin resistance gene for selection in E coli
    https://www.bioz.com/result/pentr vector/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    pentr vector - by Bioz Stars, 2021-01
    99/100 stars

    Images

    1) Product Images from "Multiple artificial microRNAs targeting conserved motifs of the replicase gene confer robust transgenic resistance to negative‐sense single‐stranded RNA plant virus"

    Article Title: Multiple artificial microRNAs targeting conserved motifs of the replicase gene confer robust transgenic resistance to negative‐sense single‐stranded RNA plant virus

    Journal: Molecular Plant Pathology

    doi: 10.1111/j.1364-3703.2011.00747.x

    Construction of artificial microRNAs (amiRNAs) targeting the conserved motifs of the tospoviral replicase sequence in the pre‐miR159a backbone. (A) Construction of single amiRNAs. MicroRNA159a (miR159a) processed from the pre‐miRNA159a backbone is presented as a hairpin. Forward primer amiR‐L‐F1 and reverse primer amiR‐L‐R, containing the complementary sequence to an individual amiRNA, were used to replace the miRNA159a and miRNA159a* sequences with an amiRNA targeting the A, B1, B2, C, D or E conserved motif of the L gene (replicase) of Watermelon silver mottle virus by first polymerase chain reaction (PCR). Bgl II‐F2 primer containing a Bgl II site and R primer containing a Sma I site were used in nested PCR. The pre‐amiRNA159a was digested with Bgl II/ Sma I, subcloned into the pENTR vector and placed downstream of the 35S promoter in a binary destination vector (destination cassette, DC) by the Gateway recombination system. (B) Strategy for triple amiRNA construction. The primer pair M13‐F/amiR159Xba‐R was used to amplify amiR‐LA (or amiR‐LB2). The primer pairs amiR159AvrII‐F/amiLB1XbaAscI‐R and amiR159AvrII‐F/amiLDXbaAscI‐R were used to amplify amiR‐LB1 and amiR‐LD, respectively. The primer pair amiR159AvrII‐F/M13‐R was used to amplify amiR‐LE or amiR‐LC. From the amplified products, the amiR‐LA or amiR‐LB2 fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LB1 or amiR‐LD fragment, respectively. After ligation, the double amiRNA segment was amplified by the primer pair M13‐F/amiLB1XbaAscI‐R or M13‐F/amiLDXbaAscI‐R. The fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LE or amiR‐LC fragment. The triple amiRNA segment was amplified by M13II‐F/pENTRAscI. The triple amiRNA fragment was released from the amplified product by Not I/ Asc I digestion and subcloned into the pENTR vector to generate pENTR‐amiR‐LAB1E and pENTR‐amiR‐LB2DC, and then into binary DC vector by Gateway recombination to form triple amiRNA constructs pre‐amiR‐LAB1E and pre‐amiR‐LB2DC.
    Figure Legend Snippet: Construction of artificial microRNAs (amiRNAs) targeting the conserved motifs of the tospoviral replicase sequence in the pre‐miR159a backbone. (A) Construction of single amiRNAs. MicroRNA159a (miR159a) processed from the pre‐miRNA159a backbone is presented as a hairpin. Forward primer amiR‐L‐F1 and reverse primer amiR‐L‐R, containing the complementary sequence to an individual amiRNA, were used to replace the miRNA159a and miRNA159a* sequences with an amiRNA targeting the A, B1, B2, C, D or E conserved motif of the L gene (replicase) of Watermelon silver mottle virus by first polymerase chain reaction (PCR). Bgl II‐F2 primer containing a Bgl II site and R primer containing a Sma I site were used in nested PCR. The pre‐amiRNA159a was digested with Bgl II/ Sma I, subcloned into the pENTR vector and placed downstream of the 35S promoter in a binary destination vector (destination cassette, DC) by the Gateway recombination system. (B) Strategy for triple amiRNA construction. The primer pair M13‐F/amiR159Xba‐R was used to amplify amiR‐LA (or amiR‐LB2). The primer pairs amiR159AvrII‐F/amiLB1XbaAscI‐R and amiR159AvrII‐F/amiLDXbaAscI‐R were used to amplify amiR‐LB1 and amiR‐LD, respectively. The primer pair amiR159AvrII‐F/M13‐R was used to amplify amiR‐LE or amiR‐LC. From the amplified products, the amiR‐LA or amiR‐LB2 fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LB1 or amiR‐LD fragment, respectively. After ligation, the double amiRNA segment was amplified by the primer pair M13‐F/amiLB1XbaAscI‐R or M13‐F/amiLDXbaAscI‐R. The fragment was digested by Xba I and ligated to Avr II‐digested amiR‐LE or amiR‐LC fragment. The triple amiRNA segment was amplified by M13II‐F/pENTRAscI. The triple amiRNA fragment was released from the amplified product by Not I/ Asc I digestion and subcloned into the pENTR vector to generate pENTR‐amiR‐LAB1E and pENTR‐amiR‐LB2DC, and then into binary DC vector by Gateway recombination to form triple amiRNA constructs pre‐amiR‐LAB1E and pre‐amiR‐LB2DC.

    Techniques Used: Sequencing, Polymerase Chain Reaction, Nested PCR, Plasmid Preparation, Amplification, Ligation, Construct

    2) Product Images from "PPARγ Regulates Genes Involved in Triacylglycerol Synthesis and Secretion in Mammary Gland Epithelial Cells of Dairy Goats"

    Article Title: PPARγ Regulates Genes Involved in Triacylglycerol Synthesis and Secretion in Mammary Gland Epithelial Cells of Dairy Goats

    Journal: PPAR Research

    doi: 10.1155/2013/310948

    Efficacy screening of the three designed shRNA via images analysis. pDsRed1-C1-PPAR γ vector was transfected as a control ((a1), (b1), and (c1)). The three tested shRNA (sh500, sh614, and sh1006) as pENTR/CMV-GFP/U6 -shRNA construct were cotransfected with pDsRed1-C1-PPAR γ vector. The transduction efficiency was estimated by the level of green fluorescent protein (GFP) expression ((a3), (b3) and (c3)). Shown are representative images of the PPAR γ expression (in red) after a 48 h cotransfection. (a1), (b1), and (c1) show high transfection and expression of PPAR γ construct vector. (a2), (b2), and (c2) show reduction of PPAR γ expression after addition of shRNA construct, while (a3), (b3), and (c3) show efficacy of shRNA transfection as shown by the green color (i.e., GFP). Images were obtained by a fluorescence microscope (Leica, DMI4000B, Germany) at 100x magnification. The images clearly show that the sh1006 had the highest effect on PPAR γ vector expression (c2).
    Figure Legend Snippet: Efficacy screening of the three designed shRNA via images analysis. pDsRed1-C1-PPAR γ vector was transfected as a control ((a1), (b1), and (c1)). The three tested shRNA (sh500, sh614, and sh1006) as pENTR/CMV-GFP/U6 -shRNA construct were cotransfected with pDsRed1-C1-PPAR γ vector. The transduction efficiency was estimated by the level of green fluorescent protein (GFP) expression ((a3), (b3) and (c3)). Shown are representative images of the PPAR γ expression (in red) after a 48 h cotransfection. (a1), (b1), and (c1) show high transfection and expression of PPAR γ construct vector. (a2), (b2), and (c2) show reduction of PPAR γ expression after addition of shRNA construct, while (a3), (b3), and (c3) show efficacy of shRNA transfection as shown by the green color (i.e., GFP). Images were obtained by a fluorescence microscope (Leica, DMI4000B, Germany) at 100x magnification. The images clearly show that the sh1006 had the highest effect on PPAR γ vector expression (c2).

    Techniques Used: shRNA, Plasmid Preparation, Transfection, Construct, Transduction, Expressing, Cotransfection, Fluorescence, Microscopy

    Related Articles

    Clone Assay:

    Article Title: Sfr1, a Tetrahymena thermophila Sfi1 Repeat Protein, Modulates the Production of Cortical Row Basal Bodies
    Article Snippet: .. The exogenous N-terminal GFP fusion used to localize Sfr1 was generated by cloning SFR1 first into the pENTR4 Dual Selection Vector (Invitrogen) and then subcloning it into pBS-MTT1 -GFP-gtw ( ) by utilizing the Gateway cloning system (Invitrogen). ..

    Article Title: Quantitative analysis of the tomato nuclear proteome during Phytophthora capsici infection unveils regulators of immunity
    Article Snippet: .. Forward primers used for amplification had CACC bases added to the 5′ end to allow cloning of amplicons into the Gateway entry vector pENTR‐D‐Topo (Life Technologies, Paisley, UK) following the manufacturer's instructions. ..

    Article Title: Clade I TGACG-Motif Binding Basic Leucine Zipper Transcription Factors Mediate BLADE-ON-PETIOLE-Dependent Regulation of Development 1Clade I TGACG-Motif Binding Basic Leucine Zipper Transcription Factors Mediate BLADE-ON-PETIOLE-Dependent Regulation of Development 1 [OPEN]
    Article Snippet: .. The resulting fragment was cloned into Gateway-compatible entry vector pENTR/D-TOPO (Invitrogen). .. Translational fusion with GFP was achieved by moving the TGA1pro:TGA1 insert into destination vector pMDC107 ( ) using LR Clonase (Invitrogen).

    Article Title: A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities
    Article Snippet: .. 4 μL of each PCR reaction were TOPO-cloned into the pENTR/D-TOPO Gateway Entry vector following the manufacturer’s protocol (pENTR Directional TOPO Cloning Kits, Invitrogen). .. Entry clones were then recombined into the pcDNA3.2/V5-DEST Gateway Destination vector at a 1:1 ratio using LR clonase II for 1 hour at 25°C (Gateway LR Clonase II Enzyme Mix, Invitrogen).

    Article Title: Multiple artificial microRNAs targeting conserved motifs of the replicase gene confer robust transgenic resistance to negative‐sense single‐stranded RNA plant virus
    Article Snippet: .. A 273‐bp fragment containing the entire sequence of the A. thaliana miR159a was cloned into the pENTR vector (Invitrogen, Carlsbad, CA, USA) to obtain pENTR‐pre‐miR159a ( ). .. The pre‐miR159a was modified into synthetic sequences targeting the highly conserved motifs A, B, C, D and E of WSMoV L RNA ( ) by two rounds of PCR ( ).

    Article Title: Optimization of the cry1Ah1 Sequence Enhances the Hyper-Resistance of Transgenic Poplars to Hyphantria cunea
    Article Snippet: .. Plasmid Construction and Transformation The synthetic full-length sequences of the cry1Ah1 genes were cloned into the Gateway entry vector pENTR/D-TOPO or pCR8/GW/TOPO (Invitrogen, Carlsbad, CA, United States), and transferred to the destination vector pH35GS ( ) by the LR reaction using LR Clonase II (Invitrogen). ..

    Transfection:

    Article Title: The S-nitrosylation of parkin attenuated the ubiquitination of divalent metal transporter 1 in MPP+-treated SH-SY5Y cells
    Article Snippet: .. When the cells grew to 70%–80% confluence, a pCMV6 vector that contained cDNA encoding parkin was transfected into cells1 (1 μg/well). .. The plasmid was mixed with the Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 30 min. Then serum-free medium was added for 4 h, followed by serum-containing medium.

    Selection:

    Article Title: Sfr1, a Tetrahymena thermophila Sfi1 Repeat Protein, Modulates the Production of Cortical Row Basal Bodies
    Article Snippet: .. The exogenous N-terminal GFP fusion used to localize Sfr1 was generated by cloning SFR1 first into the pENTR4 Dual Selection Vector (Invitrogen) and then subcloning it into pBS-MTT1 -GFP-gtw ( ) by utilizing the Gateway cloning system (Invitrogen). ..

    Modification:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Subcloning:

    Article Title: Sfr1, a Tetrahymena thermophila Sfi1 Repeat Protein, Modulates the Production of Cortical Row Basal Bodies
    Article Snippet: .. The exogenous N-terminal GFP fusion used to localize Sfr1 was generated by cloning SFR1 first into the pENTR4 Dual Selection Vector (Invitrogen) and then subcloning it into pBS-MTT1 -GFP-gtw ( ) by utilizing the Gateway cloning system (Invitrogen). ..

    Construct:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Polymerase Chain Reaction:

    Article Title: A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities
    Article Snippet: .. 4 μL of each PCR reaction were TOPO-cloned into the pENTR/D-TOPO Gateway Entry vector following the manufacturer’s protocol (pENTR Directional TOPO Cloning Kits, Invitrogen). .. Entry clones were then recombined into the pcDNA3.2/V5-DEST Gateway Destination vector at a 1:1 ratio using LR clonase II for 1 hour at 25°C (Gateway LR Clonase II Enzyme Mix, Invitrogen).

    Generated:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Article Title: Sfr1, a Tetrahymena thermophila Sfi1 Repeat Protein, Modulates the Production of Cortical Row Basal Bodies
    Article Snippet: .. The exogenous N-terminal GFP fusion used to localize Sfr1 was generated by cloning SFR1 first into the pENTR4 Dual Selection Vector (Invitrogen) and then subcloning it into pBS-MTT1 -GFP-gtw ( ) by utilizing the Gateway cloning system (Invitrogen). ..

    Amplification:

    Article Title: Quantitative analysis of the tomato nuclear proteome during Phytophthora capsici infection unveils regulators of immunity
    Article Snippet: .. Forward primers used for amplification had CACC bases added to the 5′ end to allow cloning of amplicons into the Gateway entry vector pENTR‐D‐Topo (Life Technologies, Paisley, UK) following the manufacturer's instructions. ..

    Infection:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Expressing:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Sequencing:

    Article Title: Multiple artificial microRNAs targeting conserved motifs of the replicase gene confer robust transgenic resistance to negative‐sense single‐stranded RNA plant virus
    Article Snippet: .. A 273‐bp fragment containing the entire sequence of the A. thaliana miR159a was cloned into the pENTR vector (Invitrogen, Carlsbad, CA, USA) to obtain pENTR‐pre‐miR159a ( ). .. The pre‐miR159a was modified into synthetic sequences targeting the highly conserved motifs A, B, C, D and E of WSMoV L RNA ( ) by two rounds of PCR ( ).

    Transformation Assay:

    Article Title: Optimization of the cry1Ah1 Sequence Enhances the Hyper-Resistance of Transgenic Poplars to Hyphantria cunea
    Article Snippet: .. Plasmid Construction and Transformation The synthetic full-length sequences of the cry1Ah1 genes were cloned into the Gateway entry vector pENTR/D-TOPO or pCR8/GW/TOPO (Invitrogen, Carlsbad, CA, United States), and transferred to the destination vector pH35GS ( ) by the LR reaction using LR Clonase II (Invitrogen). ..

    Plasmid Preparation:

    Article Title: Silencing collapsin response mediator protein-2 reprograms macrophage phenotype and improves infarct healing in experimental myocardial infarction model
    Article Snippet: .. Plasmids and adenoviral infection IRF expression construct was generated in the pENTR vector (Invitrogen) modified to contain the CMV promoter and IRES-linked GFP (pBent) as previously described [ ]. .. For delivery into mouse bone marrow-derived macrophages, IRF5/luciferase cassettes were excised and subcloned into the pBent vector, modified to contain CMV-driven GFP in the orientation opposite to the luciferase gene and recombined into pAD/PL DEST vector (Invitrogen) for adenovirus production.

    Article Title: Sfr1, a Tetrahymena thermophila Sfi1 Repeat Protein, Modulates the Production of Cortical Row Basal Bodies
    Article Snippet: .. The exogenous N-terminal GFP fusion used to localize Sfr1 was generated by cloning SFR1 first into the pENTR4 Dual Selection Vector (Invitrogen) and then subcloning it into pBS-MTT1 -GFP-gtw ( ) by utilizing the Gateway cloning system (Invitrogen). ..

    Article Title: The S-nitrosylation of parkin attenuated the ubiquitination of divalent metal transporter 1 in MPP+-treated SH-SY5Y cells
    Article Snippet: .. When the cells grew to 70%–80% confluence, a pCMV6 vector that contained cDNA encoding parkin was transfected into cells1 (1 μg/well). .. The plasmid was mixed with the Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA) for 30 min. Then serum-free medium was added for 4 h, followed by serum-containing medium.

    Article Title: Quantitative analysis of the tomato nuclear proteome during Phytophthora capsici infection unveils regulators of immunity
    Article Snippet: .. Forward primers used for amplification had CACC bases added to the 5′ end to allow cloning of amplicons into the Gateway entry vector pENTR‐D‐Topo (Life Technologies, Paisley, UK) following the manufacturer's instructions. ..

    Article Title: Clade I TGACG-Motif Binding Basic Leucine Zipper Transcription Factors Mediate BLADE-ON-PETIOLE-Dependent Regulation of Development 1Clade I TGACG-Motif Binding Basic Leucine Zipper Transcription Factors Mediate BLADE-ON-PETIOLE-Dependent Regulation of Development 1 [OPEN]
    Article Snippet: .. The resulting fragment was cloned into Gateway-compatible entry vector pENTR/D-TOPO (Invitrogen). .. Translational fusion with GFP was achieved by moving the TGA1pro:TGA1 insert into destination vector pMDC107 ( ) using LR Clonase (Invitrogen).

    Article Title: A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities
    Article Snippet: .. 4 μL of each PCR reaction were TOPO-cloned into the pENTR/D-TOPO Gateway Entry vector following the manufacturer’s protocol (pENTR Directional TOPO Cloning Kits, Invitrogen). .. Entry clones were then recombined into the pcDNA3.2/V5-DEST Gateway Destination vector at a 1:1 ratio using LR clonase II for 1 hour at 25°C (Gateway LR Clonase II Enzyme Mix, Invitrogen).

    Article Title: Multiple artificial microRNAs targeting conserved motifs of the replicase gene confer robust transgenic resistance to negative‐sense single‐stranded RNA plant virus
    Article Snippet: .. A 273‐bp fragment containing the entire sequence of the A. thaliana miR159a was cloned into the pENTR vector (Invitrogen, Carlsbad, CA, USA) to obtain pENTR‐pre‐miR159a ( ). .. The pre‐miR159a was modified into synthetic sequences targeting the highly conserved motifs A, B, C, D and E of WSMoV L RNA ( ) by two rounds of PCR ( ).

    Article Title: Optimization of the cry1Ah1 Sequence Enhances the Hyper-Resistance of Transgenic Poplars to Hyphantria cunea
    Article Snippet: .. Plasmid Construction and Transformation The synthetic full-length sequences of the cry1Ah1 genes were cloned into the Gateway entry vector pENTR/D-TOPO or pCR8/GW/TOPO (Invitrogen, Carlsbad, CA, United States), and transferred to the destination vector pH35GS ( ) by the LR reaction using LR Clonase II (Invitrogen). ..

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  • 99
    Thermo Fisher pcmv6 entry vector
    Increased menin expression decreases proliferation. Mz-ChA-1 cells overexpressing menin with <t>pCMV6-MEN1</t> vector exhibit a decrease in Ki-67 proliferative marker expression. A – C: Increased menin expression in pCMV6-MEN1 Mz-ChA-1 cells by real-time PCR ( A ) and flow cytometry ( B ) decreased Ki-67 proliferative marker expression by real-time PCR ( C ). D: Decreased cell migration as measured by wound healing assay. E: Decreased cell invasion as measured by Boyden chamber assay in pCMV6-MEN1 Mz-ChA-1 cells. Data are expressed as means ± SEM performed in triplicate ( A–E ). ∗ P
    Pcmv6 Entry Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcmv6 entry vector/product/Thermo Fisher
    Average 99 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    pcmv6 entry vector - by Bioz Stars, 2021-01
    99/100 stars
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    91
    Thermo Fisher control plasmid penter
    Restoration of EZH2 or HDAC9 expression reverses the effect of <t>miR-101-3p</t> on WERI-RB-1 and Y79 cells. (A) Overexpression of EZH2 or HDAC9 in indicated cells was confirmed by performing western blotting. (B) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and <t>pEnter</t> or pEnter-EZH2. Cell viability was determined by an MTT assay. (C) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and pEnter or pEnter-HDAC9. Cell viability was determined by an MTT assay. *P
    Control Plasmid Penter, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/control plasmid penter/product/Thermo Fisher
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    control plasmid penter - by Bioz Stars, 2021-01
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    99
    Thermo Fisher 3c library
    The results of <t>3C</t> library inverse PCR showed variable sizes of captured fragments due to chimeric products in 3C library preparation (A). Sample NO 8 was selected for translocation PCR primer design which has the acquirable intra-chromosomal interaction. There were no specific DNA bands obtained from genomic DNA Inverse PCR (B).This could be due to the long distance between the viewpoint locus and juxtaposed sequences in the genomic DNA.
    3c Library, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/3c library/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    3c library - by Bioz Stars, 2021-01
    99/100 stars
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    Image Search Results


    Increased menin expression decreases proliferation. Mz-ChA-1 cells overexpressing menin with pCMV6-MEN1 vector exhibit a decrease in Ki-67 proliferative marker expression. A – C: Increased menin expression in pCMV6-MEN1 Mz-ChA-1 cells by real-time PCR ( A ) and flow cytometry ( B ) decreased Ki-67 proliferative marker expression by real-time PCR ( C ). D: Decreased cell migration as measured by wound healing assay. E: Decreased cell invasion as measured by Boyden chamber assay in pCMV6-MEN1 Mz-ChA-1 cells. Data are expressed as means ± SEM performed in triplicate ( A–E ). ∗ P

    Journal: The American Journal of Pathology

    Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

    doi: 10.1016/j.ajpath.2016.10.021

    Figure Lengend Snippet: Increased menin expression decreases proliferation. Mz-ChA-1 cells overexpressing menin with pCMV6-MEN1 vector exhibit a decrease in Ki-67 proliferative marker expression. A – C: Increased menin expression in pCMV6-MEN1 Mz-ChA-1 cells by real-time PCR ( A ) and flow cytometry ( B ) decreased Ki-67 proliferative marker expression by real-time PCR ( C ). D: Decreased cell migration as measured by wound healing assay. E: Decreased cell invasion as measured by Boyden chamber assay in pCMV6-MEN1 Mz-ChA-1 cells. Data are expressed as means ± SEM performed in triplicate ( A–E ). ∗ P

    Article Snippet: The pCMV6-Entry vector without the Men1 insert was used as a control.

    Techniques: Expressing, Plasmid Preparation, Marker, Real-time Polymerase Chain Reaction, Flow Cytometry, Migration, Wound Healing Assay, Boyden Chamber Assay

    Menin expression negatively regulates angiogenesis. A: By real-time PCR, Mz-ChA-1 MEN1 knockout cells increased expression of angiogenic factors compared to Mz-ChA-1 control cells. B: By real-time PCR, pCMV6-MEN1 Mz-ChA-1 cells decreased expression of angiogenic factors compared to Mz-ChA-1 control cells. Data are expressed as means ± SEM performed in triplicate ( A and B ). ∗ P

    Journal: The American Journal of Pathology

    Article Title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation

    doi: 10.1016/j.ajpath.2016.10.021

    Figure Lengend Snippet: Menin expression negatively regulates angiogenesis. A: By real-time PCR, Mz-ChA-1 MEN1 knockout cells increased expression of angiogenic factors compared to Mz-ChA-1 control cells. B: By real-time PCR, pCMV6-MEN1 Mz-ChA-1 cells decreased expression of angiogenic factors compared to Mz-ChA-1 control cells. Data are expressed as means ± SEM performed in triplicate ( A and B ). ∗ P

    Article Snippet: The pCMV6-Entry vector without the Men1 insert was used as a control.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Knock-Out

    Restoration of EZH2 or HDAC9 expression reverses the effect of miR-101-3p on WERI-RB-1 and Y79 cells. (A) Overexpression of EZH2 or HDAC9 in indicated cells was confirmed by performing western blotting. (B) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and pEnter or pEnter-EZH2. Cell viability was determined by an MTT assay. (C) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and pEnter or pEnter-HDAC9. Cell viability was determined by an MTT assay. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: MicroRNA-101-3p inhibits proliferation in retinoblastoma cells by targeting EZH2 and HDAC9

    doi: 10.3892/etm.2018.6405

    Figure Lengend Snippet: Restoration of EZH2 or HDAC9 expression reverses the effect of miR-101-3p on WERI-RB-1 and Y79 cells. (A) Overexpression of EZH2 or HDAC9 in indicated cells was confirmed by performing western blotting. (B) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and pEnter or pEnter-EZH2. Cell viability was determined by an MTT assay. (C) WERI-RB-1 and Y79 cells were transfected with miR-control or miR-101-3p, and pEnter or pEnter-HDAC9. Cell viability was determined by an MTT assay. *P

    Article Snippet: To determine the roles of target genes on the effect of miR-101-3p, co-transfection of 50 nM miR-101-3p agomir+1 µg control plasmid (pEnter) or 50 nM miR-101-3p agomir+1 µg pEnter-EZH2/pEnter-HDAC9 plasmid in WERI-Rb-1 and Y79 cells was performed using Lipofectamine 3000 (Thermo Fisher Scientific, Inc.).

    Techniques: Expressing, Over Expression, Western Blot, Transfection, MTT Assay

    The results of 3C library inverse PCR showed variable sizes of captured fragments due to chimeric products in 3C library preparation (A). Sample NO 8 was selected for translocation PCR primer design which has the acquirable intra-chromosomal interaction. There were no specific DNA bands obtained from genomic DNA Inverse PCR (B).This could be due to the long distance between the viewpoint locus and juxtaposed sequences in the genomic DNA.

    Journal: International Journal of Hematology-Oncology and Stem Cell Research

    Article Title: Application of Chromosome Conformation Capture Method for Detection MYC/TRD Chromosomal Translocation in Leukemia Cell Line

    doi: 10.18502/ijhoscr.v14i3.3729

    Figure Lengend Snippet: The results of 3C library inverse PCR showed variable sizes of captured fragments due to chimeric products in 3C library preparation (A). Sample NO 8 was selected for translocation PCR primer design which has the acquirable intra-chromosomal interaction. There were no specific DNA bands obtained from genomic DNA Inverse PCR (B).This could be due to the long distance between the viewpoint locus and juxtaposed sequences in the genomic DNA.

    Article Snippet: The same concentration of 3C library and genomic DNA from SKW3 cell line were used for the PCR assay.

    Techniques: Inverse PCR, Translocation Assay, Polymerase Chain Reaction

    Translocation PCR performed on both 3C library and genomic DNA of SKW3 cell line. Sample 1 and 2 were indicated as 3C library and sample 3 and 4 were indicated as Digested/Ligated genomic DNA. The 184 bp (tra4) band from juxtaposed MYC /TRD was amplified in both 3C library and DNA using specific primers.

    Journal: International Journal of Hematology-Oncology and Stem Cell Research

    Article Title: Application of Chromosome Conformation Capture Method for Detection MYC/TRD Chromosomal Translocation in Leukemia Cell Line

    doi: 10.18502/ijhoscr.v14i3.3729

    Figure Lengend Snippet: Translocation PCR performed on both 3C library and genomic DNA of SKW3 cell line. Sample 1 and 2 were indicated as 3C library and sample 3 and 4 were indicated as Digested/Ligated genomic DNA. The 184 bp (tra4) band from juxtaposed MYC /TRD was amplified in both 3C library and DNA using specific primers.

    Article Snippet: The same concentration of 3C library and genomic DNA from SKW3 cell line were used for the PCR assay.

    Techniques: Translocation Assay, Polymerase Chain Reaction, Amplification

    Pericentromeric heterochromatin is organized into domains  a  schematic of  Drosophila melanogaster  chromosomes showing the pericentromeric regions (light blue) included in the 5C experiment. The centromere is in green and the euchromatic regions in dark blue.  b  Schematic representation of 5C primer design by alternating primer design scheme, at the adjacent restriction enzyme sites. Forward (with T7 overhang) and reverse (with T3 overhang) primers are used to amplify the interacting DNA junctions from the 3C library.  c  A pairwise interaction map of Chr2L showing the mapped active Het TADs green and inactive Het TADs in red, respectively, along with the epigenetic marks of H3K9me3, HP1a, H3K4me3 and H3K36me3. Active genes in the pericentromeres are enriched for H3K9me3 along with active histone modifications. MARs are indicated by red bars. TAD boundaries (grey bars) partition distinct epigenomic domains

    Journal: Epigenetics & Chromatin

    Article Title: Interplay of pericentromeric genome organization and chromatin landscape regulates the expression of Drosophila melanogaster heterochromatic genes

    doi: 10.1186/s13072-020-00358-4

    Figure Lengend Snippet: Pericentromeric heterochromatin is organized into domains a schematic of Drosophila melanogaster chromosomes showing the pericentromeric regions (light blue) included in the 5C experiment. The centromere is in green and the euchromatic regions in dark blue. b Schematic representation of 5C primer design by alternating primer design scheme, at the adjacent restriction enzyme sites. Forward (with T7 overhang) and reverse (with T3 overhang) primers are used to amplify the interacting DNA junctions from the 3C library. c A pairwise interaction map of Chr2L showing the mapped active Het TADs green and inactive Het TADs in red, respectively, along with the epigenetic marks of H3K9me3, HP1a, H3K4me3 and H3K36me3. Active genes in the pericentromeres are enriched for H3K9me3 along with active histone modifications. MARs are indicated by red bars. TAD boundaries (grey bars) partition distinct epigenomic domains

    Article Snippet: Generation of the 3C library: 5 × 107 S2 cells were grown to confluence and 95–98% viability in Schneider’s media (n = 3 biological replicates).

    Techniques: