pdonr 221 p1 p5r gateway vector  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pdonr 221 p1 p5r gateway vector
    Pdonr 221 P1 P5r Gateway Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pdonr 221 p1 p5r gateway vector/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pdonr 221 p1 p5r gateway vector - by Bioz Stars, 2020-04
    86/100 stars

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    Clone Assay:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: .. The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The PCR product of the Fred-encoding gene was recombined with the pDONR 221 P5-P2 vector to form entry clone pDONR(Fred07). pDONR(2,5-DKCMO-1) and pDONR(Fred07) were recombined with the pSD80 destination vector, which was constructed using the Gateway vector conversion system (Invitrogen) via ligation of the RfA cassette (provided in the kit) with vector pSD80, which had been linearized with the restriction enzyme BamHI and blunt ended with T4 DNA polymerase.

    Amplification:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: The template for amplification was clone pET17b-Fred as described in the previous section. .. The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO).

    Ligation:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The PCR product of the Fred-encoding gene was recombined with the pDONR 221 P5-P2 vector to form entry clone pDONR(Fred07). pDONR(2,5-DKCMO-1) and pDONR(Fred07) were recombined with the pSD80 destination vector, which was constructed using the Gateway vector conversion system (Invitrogen) via ligation of the RfA cassette (provided in the kit) with vector pSD80, which had been linearized with the restriction enzyme BamHI and blunt ended with T4 DNA polymerase.

    Construct:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: All tandem clones were constructed using a MultiSite Gateway Pro kit (Invitrogen). .. The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO).

    Expressing:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The plasmid containing the double clone (pSD80-CamE25–1 -Fred) was then transformed into E. coli BL21(DE3) for protein expression.

    Polymerase Chain Reaction:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: .. The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The PCR product of the Fred-encoding gene was recombined with the pDONR 221 P5-P2 vector to form entry clone pDONR(Fred07). pDONR(2,5-DKCMO-1) and pDONR(Fred07) were recombined with the pSD80 destination vector, which was constructed using the Gateway vector conversion system (Invitrogen) via ligation of the RfA cassette (provided in the kit) with vector pSD80, which had been linearized with the restriction enzyme BamHI and blunt ended with T4 DNA polymerase.

    Transformation Assay:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The recombination product of the three plasmids was transformed into One Shot Mach1 T1 chemically competent E. coli .

    Plasmid Preparation:

    Article Title: Camphor Pathway Redux: Functional Recombinant Expression of 2,5- and 3,6-Diketocamphane Monooxygenases of Pseudomonas putida ATCC 17453 with Their Cognate Flavin Reductase Catalyzing Baeyer-Villiger Reactions
    Article Snippet: .. The various camE PCR products were recombined with the pDONR 221 P1-P5r Gateway vector (Invitrogen) in separate reactions to form entry clones pDONR(2,5-DKCMO-1), pDONR(2,5-DKCMO-2), and pDONR(3,6-DKCMO). .. The PCR product of the Fred-encoding gene was recombined with the pDONR 221 P5-P2 vector to form entry clone pDONR(Fred07). pDONR(2,5-DKCMO-1) and pDONR(Fred07) were recombined with the pSD80 destination vector, which was constructed using the Gateway vector conversion system (Invitrogen) via ligation of the RfA cassette (provided in the kit) with vector pSD80, which had been linearized with the restriction enzyme BamHI and blunt ended with T4 DNA polymerase.

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  • 93
    Thermo Fisher pdonr 221 p1 p5r
    Pdonr 221 P1 P5r, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pdonr 221 p1 p5r/product/Thermo Fisher
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    pdonr 221 p1 p5r - by Bioz Stars, 2020-04
    93/100 stars
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