chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    a , Domain organization of the carbohydrate diacid regulator CdaR. Diacid binding domain and shared chemotypes between glycerate, D-glucarate, and D-galactarate are shown in blue. b , Testing CdaR-regulated promoters using glycerate and glycolate, nominating the glycerate-specific P garP with the greatest signal and dynamic range. c , Schematic representation of the optimized glycerate-responsive biosensor: CdaR is constitutively expressed and requires glycerate (GLY) to activate P garP transcription, resulting in a luminescent signal through LuxAB. Grayscale elements correspond to the origin of replication and resistance marker. d , The single plasmid glycerate-responsive biosensor improves dynamic range in S4221 cells, but is constitutively activated in the evolved strains S4009 and S4012, suggesting unintended crosstalk with an ALE-derived cellular metabolite. e , Selection strategy to improve CdaR substrate specificity in S4012 cells: negative selection (without glycerate): undesired promoter activation produces the toxin barnase and results in cell death. Positive selection (with glycerate): barnase is inhibited by the constitutively expressed antitoxin barstar, and glycerate-dependent activation produces a fluorescent signal (mCherry) and <t>chloramphenicol</t> resistance (Cat). f , Selection outcomes depend on the medium used for evolution (evo) and subsequent assays (test). Directed evolution of CdaR in DRM yielded variants with greatest biosensor activation. The labels “evo” and “test” indicate the medium used for directed evolution and the medium used in the shown assay, respectively. g , Single clone analysis and Sanger sequencing of variants with large dynamic range yielded two genotypes: Cdar evo1 and Cdar evo2 . h , Mutational mapping of evolved mutations onto an Alphafold-predicted , CdaR dimer shows that mutations may affect a putative ligand binding pocket at the dimer interface. Glycerate docked using CB-Dock , . Evolved mutations are shown schematically in the CdaR diacid binding domain. Red (Cdar evo1 ) and orange (Cdar evo2 ) mutations are coding, whereas mutations in white are non-coding. i , Cdar evo1 restores glycerate-dependent biosensor activity in the metabolically insulated strains S4009 and S4012. j , Cdar evo1 benchmarking using various strains and reporters. S4012 cells encoding Cdar evo1 driving an mScarlet-I reporter were used for all subsequent assays. All datasets include 4 biological replicates.
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A Genetically Encoded System to Quantify and Evolve RuBisCO-Catalyzed Carbon Fixation"

    Article Title: A Genetically Encoded System to Quantify and Evolve RuBisCO-Catalyzed Carbon Fixation

    Journal: bioRxiv

    doi: 10.1101/2024.05.15.594374

    a , Domain organization of the carbohydrate diacid regulator CdaR. Diacid binding domain and shared chemotypes between glycerate, D-glucarate, and D-galactarate are shown in blue. b , Testing CdaR-regulated promoters using glycerate and glycolate, nominating the glycerate-specific P garP with the greatest signal and dynamic range. c , Schematic representation of the optimized glycerate-responsive biosensor: CdaR is constitutively expressed and requires glycerate (GLY) to activate P garP transcription, resulting in a luminescent signal through LuxAB. Grayscale elements correspond to the origin of replication and resistance marker. d , The single plasmid glycerate-responsive biosensor improves dynamic range in S4221 cells, but is constitutively activated in the evolved strains S4009 and S4012, suggesting unintended crosstalk with an ALE-derived cellular metabolite. e , Selection strategy to improve CdaR substrate specificity in S4012 cells: negative selection (without glycerate): undesired promoter activation produces the toxin barnase and results in cell death. Positive selection (with glycerate): barnase is inhibited by the constitutively expressed antitoxin barstar, and glycerate-dependent activation produces a fluorescent signal (mCherry) and chloramphenicol resistance (Cat). f , Selection outcomes depend on the medium used for evolution (evo) and subsequent assays (test). Directed evolution of CdaR in DRM yielded variants with greatest biosensor activation. The labels “evo” and “test” indicate the medium used for directed evolution and the medium used in the shown assay, respectively. g , Single clone analysis and Sanger sequencing of variants with large dynamic range yielded two genotypes: Cdar evo1 and Cdar evo2 . h , Mutational mapping of evolved mutations onto an Alphafold-predicted , CdaR dimer shows that mutations may affect a putative ligand binding pocket at the dimer interface. Glycerate docked using CB-Dock , . Evolved mutations are shown schematically in the CdaR diacid binding domain. Red (Cdar evo1 ) and orange (Cdar evo2 ) mutations are coding, whereas mutations in white are non-coding. i , Cdar evo1 restores glycerate-dependent biosensor activity in the metabolically insulated strains S4009 and S4012. j , Cdar evo1 benchmarking using various strains and reporters. S4012 cells encoding Cdar evo1 driving an mScarlet-I reporter were used for all subsequent assays. All datasets include 4 biological replicates.
    Figure Legend Snippet: a , Domain organization of the carbohydrate diacid regulator CdaR. Diacid binding domain and shared chemotypes between glycerate, D-glucarate, and D-galactarate are shown in blue. b , Testing CdaR-regulated promoters using glycerate and glycolate, nominating the glycerate-specific P garP with the greatest signal and dynamic range. c , Schematic representation of the optimized glycerate-responsive biosensor: CdaR is constitutively expressed and requires glycerate (GLY) to activate P garP transcription, resulting in a luminescent signal through LuxAB. Grayscale elements correspond to the origin of replication and resistance marker. d , The single plasmid glycerate-responsive biosensor improves dynamic range in S4221 cells, but is constitutively activated in the evolved strains S4009 and S4012, suggesting unintended crosstalk with an ALE-derived cellular metabolite. e , Selection strategy to improve CdaR substrate specificity in S4012 cells: negative selection (without glycerate): undesired promoter activation produces the toxin barnase and results in cell death. Positive selection (with glycerate): barnase is inhibited by the constitutively expressed antitoxin barstar, and glycerate-dependent activation produces a fluorescent signal (mCherry) and chloramphenicol resistance (Cat). f , Selection outcomes depend on the medium used for evolution (evo) and subsequent assays (test). Directed evolution of CdaR in DRM yielded variants with greatest biosensor activation. The labels “evo” and “test” indicate the medium used for directed evolution and the medium used in the shown assay, respectively. g , Single clone analysis and Sanger sequencing of variants with large dynamic range yielded two genotypes: Cdar evo1 and Cdar evo2 . h , Mutational mapping of evolved mutations onto an Alphafold-predicted , CdaR dimer shows that mutations may affect a putative ligand binding pocket at the dimer interface. Glycerate docked using CB-Dock , . Evolved mutations are shown schematically in the CdaR diacid binding domain. Red (Cdar evo1 ) and orange (Cdar evo2 ) mutations are coding, whereas mutations in white are non-coding. i , Cdar evo1 restores glycerate-dependent biosensor activity in the metabolically insulated strains S4009 and S4012. j , Cdar evo1 benchmarking using various strains and reporters. S4012 cells encoding Cdar evo1 driving an mScarlet-I reporter were used for all subsequent assays. All datasets include 4 biological replicates.

    Techniques Used: Binding Assay, Marker, Plasmid Preparation, Derivative Assay, Selection, Activation Assay, Sequencing, Ligand Binding Assay, Activity Assay, Metabolic Labelling

    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
    Chloramphenicol, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
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    chloramphenicol  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol
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    chloramphenicol cam  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol cam
    The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by <t>chloramphenicol</t> resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).
    Chloramphenicol Cam, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Methionyl-tRNA synthetase synthetic and proofreading activities are determinants of antibiotic persistence"

    Article Title: Methionyl-tRNA synthetase synthetic and proofreading activities are determinants of antibiotic persistence

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2024.1384552

    The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by chloramphenicol resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).
    Figure Legend Snippet: The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by chloramphenicol resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).

    Techniques Used: Plasmid Preparation, Sequencing, Conjugation Assay, Construct

    chloramphenicol goldbio c 105 25 25  (Gold Biotechnology Inc)


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    Gold Biotechnology Inc chloramphenicol goldbio c 105 25 25
    Chloramphenicol Goldbio C 105 25 25, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gold Biotechnology Inc chloramphenicol
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    Gold Biotechnology Inc chloramphenicol cam
    The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by <t>chloramphenicol</t> resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).
    Chloramphenicol Cam, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gold Biotechnology Inc chloramphenicol goldbio c 105 25 25
    The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by <t>chloramphenicol</t> resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).
    Chloramphenicol Goldbio C 105 25 25, supplied by Gold Biotechnology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by chloramphenicol resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).

    Journal: Frontiers in Microbiology

    Article Title: Methionyl-tRNA synthetase synthetic and proofreading activities are determinants of antibiotic persistence

    doi: 10.3389/fmicb.2024.1384552

    Figure Lengend Snippet: The R6K integrated-plasmid recombination (RIPR) system for genome editing by allelic exchange. (A) The pRIPR vector contains π dependent R6K origin of replication ( oriR6K ), origin of transfer oriT RP4 and the 18-bp I-SceI meganuclease recognition site. Restriction enzyme recognition sites (e.g., BspQI) facilitate insertion of DNA containing the desired sequence change (e.g., GAA, encoding glutamate [Glu]) flanked by homology arms (YFG). Plasmid transformants and recombinants are selected by chloramphenicol resistance ( cam ). (B) Following conjugation into a recipient cell, pRIPR integrates into the bacterial chromosome by a single-crossover event. Production of I-SceI from the helper plasmid pSceH (not shown) generates a double-stranded DNA break as a counterselection against plasmid integrates. (C) Resolution of the cointegrate by a second recombination event results in the loss of RIPR construct leaving behind the desired sequence change (GAA) or the wild type sequence (GCA).

    Article Snippet: Bacteria were grown using Miller Broth Base (Fisher Scientific) and supplemented with antibiotics ampicillin (Amp) (100 μg/mL), chloramphenicol (Cam) (30 μg/mL) and kanamycin (Kan) (30 μg/mL), (GoldBio).

    Techniques: Plasmid Preparation, Sequencing, Conjugation Assay, Construct