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  • 99
    Thermo Fisher gateway lr clonase ii enzyme mix
    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite <t>Clonase</t> strategy utilizing modified MuLE entry vectors.
    Gateway Lr Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7014 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher lr clonase ii enzyme mix
    The rationale of BioVector. The LR reaction among GEC, PEC and BDV shows the basic principle of BioVector, in which a gene and a promoter in individual entry clones are positioningly and directionally joined together in a destination vector. att L1/2/3/4, att R1/2/3/4, and att P1/2/3/4, recombination sites; ccd B, a negative selection marker for DH5α; LR <t>Clonase</t> II®, recombination enzyme from Invitrogen.
    Lr Clonase Ii Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3247 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase ii enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 3247 article reviews
    Price from $9.99 to $1999.99
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    99
    Thermo Fisher lr clonase enzyme mix
    Engineering the CRISPR-Cas9 machinery for interference against TYLCV . The T-DNAs, U6-sgRNA and Cas9, were cloned into the binary vector pK2GW7 by restriction ligation and Gateway LR <t>clonase,</t> respectively. Agrobacterium containing the engineered T-DNA vector for targeting the TYLCV genome was used to transform tomato ( S. lycopersicum ) and N. benthamiana plants. LB, left border. Kan R , plant selection kanamycin gene. U6, U6-26 Arabidopsis promoter. p35S, CaMV-35S promoter. 3x-flag, flag tag for confirmation of protein expression. NLS, nuclear localization signal. pCas9, human codon-optimized Cas9 of S. pyogenes . RB, right border. SmR, spectinomycin bacterial selection. B-D) T7EI assay for TYLCV targeting at the Rep sequence (B), the CP sequence (C), and the Ssp I restriction enzyme site for the IR sequence (D) in three independent T2 lines of N. benthamiana . A PCR amplified fragment flanking the targets in the CP and Rep sequence of the TYLCV genome were subjected to the T7EI assay to detect InDels. InDels were detected only in PCR amplicons from plants expressing Cas9 and sgRNA, but not in the wild type used as the control. E) CRISPR-Cas9-mediated virus targeting in T 3 plants of N. benthamiana . The T7EI assay confirmed CRISPR-Cas9-mediated targeting of the TYLCV in two independent T 3 plants expressing Cas9 and sgRNA, but not in the wild type used as the control. In B, C, D, and E, DNA fragments were resolved on a 2% agarose gel and stained with ethidium bromide; the ratio of InDels are represented as a percentage below each gel, and arrows indicate the two T7EI digested fragments. F) Dot-blot analysis for the accumulation of the TYLCV genome in CRISPR/Cas9-expressing T 3 N. benthamiana plants. Total DNA extracted (10 ng) from three independent T 3 plants was blotted, probed with DIG-labeled TYLCV fragments, and detected with anti-DIG antibodies using the (Roche GmbH) protocol.
    Lr Clonase Enzyme Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1259 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase enzyme mix/product/Thermo Fisher
    Average 99 stars, based on 1259 article reviews
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    lr clonase enzyme mix - by Bioz Stars, 2020-09
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    89
    Thermo Fisher lr clonase ii enzyme
    Engineering the CRISPR-Cas9 machinery for interference against TYLCV . The T-DNAs, U6-sgRNA and Cas9, were cloned into the binary vector pK2GW7 by restriction ligation and Gateway LR <t>clonase,</t> respectively. Agrobacterium containing the engineered T-DNA vector for targeting the TYLCV genome was used to transform tomato ( S. lycopersicum ) and N. benthamiana plants. LB, left border. Kan R , plant selection kanamycin gene. U6, U6-26 Arabidopsis promoter. p35S, CaMV-35S promoter. 3x-flag, flag tag for confirmation of protein expression. NLS, nuclear localization signal. pCas9, human codon-optimized Cas9 of S. pyogenes . RB, right border. SmR, spectinomycin bacterial selection. B-D) T7EI assay for TYLCV targeting at the Rep sequence (B), the CP sequence (C), and the Ssp I restriction enzyme site for the IR sequence (D) in three independent T2 lines of N. benthamiana . A PCR amplified fragment flanking the targets in the CP and Rep sequence of the TYLCV genome were subjected to the T7EI assay to detect InDels. InDels were detected only in PCR amplicons from plants expressing Cas9 and sgRNA, but not in the wild type used as the control. E) CRISPR-Cas9-mediated virus targeting in T 3 plants of N. benthamiana . The T7EI assay confirmed CRISPR-Cas9-mediated targeting of the TYLCV in two independent T 3 plants expressing Cas9 and sgRNA, but not in the wild type used as the control. In B, C, D, and E, DNA fragments were resolved on a 2% agarose gel and stained with ethidium bromide; the ratio of InDels are represented as a percentage below each gel, and arrows indicate the two T7EI digested fragments. F) Dot-blot analysis for the accumulation of the TYLCV genome in CRISPR/Cas9-expressing T 3 N. benthamiana plants. Total DNA extracted (10 ng) from three independent T 3 plants was blotted, probed with DIG-labeled TYLCV fragments, and detected with anti-DIG antibodies using the (Roche GmbH) protocol.
    Lr Clonase Ii Enzyme, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase ii enzyme/product/Thermo Fisher
    Average 89 stars, based on 496 article reviews
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    lr clonase ii enzyme - by Bioz Stars, 2020-09
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    95
    Thermo Fisher lr clonase ii kit
    Engineering the CRISPR-Cas9 machinery for interference against TYLCV . The T-DNAs, U6-sgRNA and Cas9, were cloned into the binary vector pK2GW7 by restriction ligation and Gateway LR <t>clonase,</t> respectively. Agrobacterium containing the engineered T-DNA vector for targeting the TYLCV genome was used to transform tomato ( S. lycopersicum ) and N. benthamiana plants. LB, left border. Kan R , plant selection kanamycin gene. U6, U6-26 Arabidopsis promoter. p35S, CaMV-35S promoter. 3x-flag, flag tag for confirmation of protein expression. NLS, nuclear localization signal. pCas9, human codon-optimized Cas9 of S. pyogenes . RB, right border. SmR, spectinomycin bacterial selection. B-D) T7EI assay for TYLCV targeting at the Rep sequence (B), the CP sequence (C), and the Ssp I restriction enzyme site for the IR sequence (D) in three independent T2 lines of N. benthamiana . A PCR amplified fragment flanking the targets in the CP and Rep sequence of the TYLCV genome were subjected to the T7EI assay to detect InDels. InDels were detected only in PCR amplicons from plants expressing Cas9 and sgRNA, but not in the wild type used as the control. E) CRISPR-Cas9-mediated virus targeting in T 3 plants of N. benthamiana . The T7EI assay confirmed CRISPR-Cas9-mediated targeting of the TYLCV in two independent T 3 plants expressing Cas9 and sgRNA, but not in the wild type used as the control. In B, C, D, and E, DNA fragments were resolved on a 2% agarose gel and stained with ethidium bromide; the ratio of InDels are represented as a percentage below each gel, and arrows indicate the two T7EI digested fragments. F) Dot-blot analysis for the accumulation of the TYLCV genome in CRISPR/Cas9-expressing T 3 N. benthamiana plants. Total DNA extracted (10 ng) from three independent T 3 plants was blotted, probed with DIG-labeled TYLCV fragments, and detected with anti-DIG antibodies using the (Roche GmbH) protocol.
    Lr Clonase Ii Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 264 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lr clonase ii kit/product/Thermo Fisher
    Average 95 stars, based on 264 article reviews
    Price from $9.99 to $1999.99
    lr clonase ii kit - by Bioz Stars, 2020-09
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    Image Search Results


    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Journal: Gene

    Article Title: A Splice Junction-Targeted CRISPR Approach (spJCRISPR) Reveals Human FOXO3B To Be A Protein-Coding Gene

    doi: 10.1016/j.gene.2018.06.048

    Figure Lengend Snippet: FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Article Snippet: LR Clonase II (Thermo 11791100) reactions were performed with a modified form of the pInducer20 vector (Addgene #44012) where the Neo resistance cassette was replaced with a blasticidin resistance cassette.

    Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Modification

    The rationale of BioVector. The LR reaction among GEC, PEC and BDV shows the basic principle of BioVector, in which a gene and a promoter in individual entry clones are positioningly and directionally joined together in a destination vector. att L1/2/3/4, att R1/2/3/4, and att P1/2/3/4, recombination sites; ccd B, a negative selection marker for DH5α; LR Clonase II®, recombination enzyme from Invitrogen.

    Journal: BMC Plant Biology

    Article Title: BioVector, a flexible system for gene specific-expression in plants

    doi: 10.1186/1471-2229-13-198

    Figure Lengend Snippet: The rationale of BioVector. The LR reaction among GEC, PEC and BDV shows the basic principle of BioVector, in which a gene and a promoter in individual entry clones are positioningly and directionally joined together in a destination vector. att L1/2/3/4, att R1/2/3/4, and att P1/2/3/4, recombination sites; ccd B, a negative selection marker for DH5α; LR Clonase II®, recombination enzyme from Invitrogen.

    Article Snippet: 5 μL reaction system was used for multiple-components LR, including Fu39-2 (20 ~ 50 ng), two entry clones (120 ~ 150ng, respectively) and 1 μL LR Clonase II Enzyme mix (Invitrogen).

    Techniques: Clone Assay, Plasmid Preparation, Selection, Marker

    Subcellular localization of CpEBF1, CpMADS1/3, and CpEIL1. All of the pENTR vectors were incubated with the pGWB5 vector together with LR clonase (Invitrogen), resulting in CpEBF1-GFP, CpMADS1/3-GFP, and CpEIL1-GFP fusion proteins. Leaves from 3- to 5-week-old Nicotiana benthamiana plants were infiltrated with GV3101 strains containing the pGWB5 recombinant vector. An empty pGWB5 vector that carries free GFP was used as the control. All the fluorescence microscopy observation assays were repeated at least thrice. Bars = 50 μM

    Journal: Horticulture Research

    Article Title: The interaction of CpEBF1 with CpMADSs is involved in cell wall degradation during papaya fruit ripening

    doi: 10.1038/s41438-018-0095-1

    Figure Lengend Snippet: Subcellular localization of CpEBF1, CpMADS1/3, and CpEIL1. All of the pENTR vectors were incubated with the pGWB5 vector together with LR clonase (Invitrogen), resulting in CpEBF1-GFP, CpMADS1/3-GFP, and CpEIL1-GFP fusion proteins. Leaves from 3- to 5-week-old Nicotiana benthamiana plants were infiltrated with GV3101 strains containing the pGWB5 recombinant vector. An empty pGWB5 vector that carries free GFP was used as the control. All the fluorescence microscopy observation assays were repeated at least thrice. Bars = 50 μM

    Article Snippet: The pENTR vectors were incubated with a pGWB5 vector together with an LR clonase enzyme (Invitrogen, USA) to generate the CpEBF1-GFP, CpMADS1/3-GFP, and CpEIL1-GFP fusion proteins.

    Techniques: Incubation, Plasmid Preparation, Recombinant, Fluorescence, Microscopy

    FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Journal: Gene

    Article Title: A Splice Junction-Targeted CRISPR Approach (spJCRISPR) Reveals Human FOXO3B To Be A Protein-Coding Gene

    doi: 10.1016/j.gene.2018.06.048

    Figure Lengend Snippet: FOXO3A and FOXO3B can be targeted for deletion independently with the spJCRISPR approach. A. Cartoon depiction of spJCRISPR targeting strategy for FOXO3A and FOXO3B loci. Red indicates 5p and 3p UTRs, green intronic sequence and blue protein coding sequence. Red arrows indicate sgRNAs, black arrows primers used for both RT-PCR and genomic PCR, blue arrows primers used for genomic PCR only and green arrows RT-PCR only. B. A cartoon depiction of the multisite Clonase strategy utilizing modified MuLE entry vectors.

    Article Snippet: LR Clonase II Plus (Thermo 12538) reactions were performed with 2 μl Clonase LR II Plus, 20 fmoles pLenti X1 Puro DEST, 10 fmoles of each sgRNA pENTR and 10 fmoles of pENTR hCas9D10A all to 10 μl volume in Tris-EDTA (TE) buffer pH 8.

    Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Modification

    Strategy for the cloning of fusion proteins. Cloning is based on the Multi-site Gateway kit from Invitrogen. The plasmid encoding the ORF of interest can either be an existing entry clone from a Gateway-based ORFeome library, or can be constructed by Gateway-cloning the PCR-amplified ORF into pDONR221 with BP clonase (panel 1) . A collection of entry clones encoding the functional modules to be attached to the ORF of interest is constructed similarly by Gateway cloning the PCR-amplified modules into the pDONR P4-P1R (N-term modules) and pDONR P2RP3 (C-term modules) plasmids from the MultiSite Gateway kit ( panel 2 , see main text for details). All of the inserts in the entry vectors are flanked by appropriate att sites provided by the PCR-amplification primers, and are indicated throughout this figure with a key (e.g. the attB1 site is indicated as B1 ). Single-fragment Gateway destination expression vectors designed for expression in different model systems are adapted by engineering of the Gateway cassette to allow MultiSite Gateway cloning reactions (panel 3) . Such reactions involve one adapted destination vector, as well as the plasmids encoding the ORF of interest and the chosen N-term and C-term modules (white background), whose matching att sites (R4xL4, R1xL1, L2xR2, L3xR3) recombine to produce a destination expression vector encoding the fusion protein in the form of a chimeric cDNA (panel 4) . Expression of the cDNA results in the production of a fusion protein whose three constituent parts are linked by short peptides resulting from the translation of the attB1 and attB2 sites remaining after the LR recombination (panel 4, see main text for details).

    Journal: BMC Molecular Biology

    Article Title: A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector

    doi: 10.1186/1471-2199-14-18

    Figure Lengend Snippet: Strategy for the cloning of fusion proteins. Cloning is based on the Multi-site Gateway kit from Invitrogen. The plasmid encoding the ORF of interest can either be an existing entry clone from a Gateway-based ORFeome library, or can be constructed by Gateway-cloning the PCR-amplified ORF into pDONR221 with BP clonase (panel 1) . A collection of entry clones encoding the functional modules to be attached to the ORF of interest is constructed similarly by Gateway cloning the PCR-amplified modules into the pDONR P4-P1R (N-term modules) and pDONR P2RP3 (C-term modules) plasmids from the MultiSite Gateway kit ( panel 2 , see main text for details). All of the inserts in the entry vectors are flanked by appropriate att sites provided by the PCR-amplification primers, and are indicated throughout this figure with a key (e.g. the attB1 site is indicated as B1 ). Single-fragment Gateway destination expression vectors designed for expression in different model systems are adapted by engineering of the Gateway cassette to allow MultiSite Gateway cloning reactions (panel 3) . Such reactions involve one adapted destination vector, as well as the plasmids encoding the ORF of interest and the chosen N-term and C-term modules (white background), whose matching att sites (R4xL4, R1xL1, L2xR2, L3xR3) recombine to produce a destination expression vector encoding the fusion protein in the form of a chimeric cDNA (panel 4) . Expression of the cDNA results in the production of a fusion protein whose three constituent parts are linked by short peptides resulting from the translation of the attB1 and attB2 sites remaining after the LR recombination (panel 4, see main text for details).

    Article Snippet: BP- and LR-recombination reactions were carried out with BP clonase II and LR Clonase II Plus enzyme mixes, respectively (Life Technologies), following the manufacturer’s instructions.

    Techniques: Clone Assay, Plasmid Preparation, Construct, Polymerase Chain Reaction, Amplification, Functional Assay, Expressing

    Engineering the CRISPR-Cas9 machinery for interference against TYLCV . The T-DNAs, U6-sgRNA and Cas9, were cloned into the binary vector pK2GW7 by restriction ligation and Gateway LR clonase, respectively. Agrobacterium containing the engineered T-DNA vector for targeting the TYLCV genome was used to transform tomato ( S. lycopersicum ) and N. benthamiana plants. LB, left border. Kan R , plant selection kanamycin gene. U6, U6-26 Arabidopsis promoter. p35S, CaMV-35S promoter. 3x-flag, flag tag for confirmation of protein expression. NLS, nuclear localization signal. pCas9, human codon-optimized Cas9 of S. pyogenes . RB, right border. SmR, spectinomycin bacterial selection. B-D) T7EI assay for TYLCV targeting at the Rep sequence (B), the CP sequence (C), and the Ssp I restriction enzyme site for the IR sequence (D) in three independent T2 lines of N. benthamiana . A PCR amplified fragment flanking the targets in the CP and Rep sequence of the TYLCV genome were subjected to the T7EI assay to detect InDels. InDels were detected only in PCR amplicons from plants expressing Cas9 and sgRNA, but not in the wild type used as the control. E) CRISPR-Cas9-mediated virus targeting in T 3 plants of N. benthamiana . The T7EI assay confirmed CRISPR-Cas9-mediated targeting of the TYLCV in two independent T 3 plants expressing Cas9 and sgRNA, but not in the wild type used as the control. In B, C, D, and E, DNA fragments were resolved on a 2% agarose gel and stained with ethidium bromide; the ratio of InDels are represented as a percentage below each gel, and arrows indicate the two T7EI digested fragments. F) Dot-blot analysis for the accumulation of the TYLCV genome in CRISPR/Cas9-expressing T 3 N. benthamiana plants. Total DNA extracted (10 ng) from three independent T 3 plants was blotted, probed with DIG-labeled TYLCV fragments, and detected with anti-DIG antibodies using the (Roche GmbH) protocol.

    Journal: Plant Signaling & Behavior

    Article Title: Engineering resistance against Tomato yellow leaf curl virus via the CRISPR/Cas9 system in tomato

    doi: 10.1080/15592324.2018.1525996

    Figure Lengend Snippet: Engineering the CRISPR-Cas9 machinery for interference against TYLCV . The T-DNAs, U6-sgRNA and Cas9, were cloned into the binary vector pK2GW7 by restriction ligation and Gateway LR clonase, respectively. Agrobacterium containing the engineered T-DNA vector for targeting the TYLCV genome was used to transform tomato ( S. lycopersicum ) and N. benthamiana plants. LB, left border. Kan R , plant selection kanamycin gene. U6, U6-26 Arabidopsis promoter. p35S, CaMV-35S promoter. 3x-flag, flag tag for confirmation of protein expression. NLS, nuclear localization signal. pCas9, human codon-optimized Cas9 of S. pyogenes . RB, right border. SmR, spectinomycin bacterial selection. B-D) T7EI assay for TYLCV targeting at the Rep sequence (B), the CP sequence (C), and the Ssp I restriction enzyme site for the IR sequence (D) in three independent T2 lines of N. benthamiana . A PCR amplified fragment flanking the targets in the CP and Rep sequence of the TYLCV genome were subjected to the T7EI assay to detect InDels. InDels were detected only in PCR amplicons from plants expressing Cas9 and sgRNA, but not in the wild type used as the control. E) CRISPR-Cas9-mediated virus targeting in T 3 plants of N. benthamiana . The T7EI assay confirmed CRISPR-Cas9-mediated targeting of the TYLCV in two independent T 3 plants expressing Cas9 and sgRNA, but not in the wild type used as the control. In B, C, D, and E, DNA fragments were resolved on a 2% agarose gel and stained with ethidium bromide; the ratio of InDels are represented as a percentage below each gel, and arrows indicate the two T7EI digested fragments. F) Dot-blot analysis for the accumulation of the TYLCV genome in CRISPR/Cas9-expressing T 3 N. benthamiana plants. Total DNA extracted (10 ng) from three independent T 3 plants was blotted, probed with DIG-labeled TYLCV fragments, and detected with anti-DIG antibodies using the (Roche GmbH) protocol.

    Article Snippet: Next, the 3XFlag–NLS-Cas9-NLS cassette was cloned into the pENTR/D-TOPO plasmid (Life Technologies) and moved to the U6-sgRNA-pK2GW7 destination vector by Gateway LR clonase in front of the CaMV-35S promoter to make the U6-sgRNA-pK2-Cas9 T-DNA vector (Life Technologies).

    Techniques: CRISPR, Clone Assay, Plasmid Preparation, Ligation, Selection, FLAG-tag, Expressing, T7EI Assay, Sequencing, Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining, Dot Blot, Labeling

    Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.

    Journal: Plant Methods

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens

    doi: 10.1186/1746-4811-7-42

    Figure Lengend Snippet: Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.

    Article Snippet: LR reaction and transformation of Agrobacterium tumefaciens by electroporation The Gateway® LR clonase enzyme mix kit (Invitrogen) was used for the LR recombination reaction.

    Techniques: Clone Assay, Expressing, Plasmid Preparation, Selection