t7 express lysy iq cells  (New England Biolabs)


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    t7 express lysy iq cells  (New England Biolabs)


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    e coli bl21 t7 express lysy iq cells  (New England Biolabs)


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    c3013 cells  (New England Biolabs)


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    e coli t7 express lysy iq competent cells  (New England Biolabs)


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    e coli t7 express lysy iq competent cells  (New England Biolabs)


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    New England Biolabs e coli t7 express lysy iq competent cells
    a , Schematic of the Clostridium symbiosum and Clostridioides difficile bilRS construct. P tac is a tac-promoter, P <t>T7</t> is a T7 promoter, and LacO is a lac operator. b , Schematic of the Ruminococcus gnavus bilR construct. c , Fluorescence assay comparing bilirubin reduction activities of <t>E.</t> <t>coli</t> 10-beta transformed with the empty vector, Clostridium symbiosum construct and Clostridioides difficile construct (two-sample one-sided t -test). d , Metabolomics confirmation of urobilin production (two-sample one-sided t -test). e , Fluorescence confirmation of the bilirubin reduction activity of E. coli 10-beta transformed with the Ruminococcus gnavus bilR construct (two-sample t -test). The bar heights show the mean of n = 4 independent biological replicates used to generate the plots and statistics in c and d . A total of n = 3 independent biological replicates were used to generate the plots and statistics in e . Error bars indicate 1 s.e. above and below the mean. P values of a two-sample one-sided t -test to determine whether the mean of the samples is greater than the mean of the vector controls are provided above the brackets. Individual data points are indicated by black points on each plot.
    E Coli T7 Express Lysy Iq Competent Cells, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "BilR is a gut microbial enzyme that reduces bilirubin to urobilinogen"

    Article Title: BilR is a gut microbial enzyme that reduces bilirubin to urobilinogen

    Journal: Nature Microbiology

    doi: 10.1038/s41564-023-01549-x

    a , Schematic of the Clostridium symbiosum and Clostridioides difficile bilRS construct. P tac is a tac-promoter, P T7 is a T7 promoter, and LacO is a lac operator. b , Schematic of the Ruminococcus gnavus bilR construct. c , Fluorescence assay comparing bilirubin reduction activities of E. coli 10-beta transformed with the empty vector, Clostridium symbiosum construct and Clostridioides difficile construct (two-sample one-sided t -test). d , Metabolomics confirmation of urobilin production (two-sample one-sided t -test). e , Fluorescence confirmation of the bilirubin reduction activity of E. coli 10-beta transformed with the Ruminococcus gnavus bilR construct (two-sample t -test). The bar heights show the mean of n = 4 independent biological replicates used to generate the plots and statistics in c and d . A total of n = 3 independent biological replicates were used to generate the plots and statistics in e . Error bars indicate 1 s.e. above and below the mean. P values of a two-sample one-sided t -test to determine whether the mean of the samples is greater than the mean of the vector controls are provided above the brackets. Individual data points are indicated by black points on each plot.
    Figure Legend Snippet: a , Schematic of the Clostridium symbiosum and Clostridioides difficile bilRS construct. P tac is a tac-promoter, P T7 is a T7 promoter, and LacO is a lac operator. b , Schematic of the Ruminococcus gnavus bilR construct. c , Fluorescence assay comparing bilirubin reduction activities of E. coli 10-beta transformed with the empty vector, Clostridium symbiosum construct and Clostridioides difficile construct (two-sample one-sided t -test). d , Metabolomics confirmation of urobilin production (two-sample one-sided t -test). e , Fluorescence confirmation of the bilirubin reduction activity of E. coli 10-beta transformed with the Ruminococcus gnavus bilR construct (two-sample t -test). The bar heights show the mean of n = 4 independent biological replicates used to generate the plots and statistics in c and d . A total of n = 3 independent biological replicates were used to generate the plots and statistics in e . Error bars indicate 1 s.e. above and below the mean. P values of a two-sample one-sided t -test to determine whether the mean of the samples is greater than the mean of the vector controls are provided above the brackets. Individual data points are indicated by black points on each plot.

    Techniques Used: Construct, Fluorescence, Transformation Assay, Plasmid Preparation, Activity Assay

    a) Fluorescence assay comparing mesobilirubin reduction activity of C. difficile CD3, C. symbiosum WAL-14163, and R. gnavus to an abiotic media controls with mesobilirubin added. b) Fluorescence assay comparing mesobilirubin reduction activity of heterologously expressed C. symbiosum bilRS in E. coli to the vector control (two-sample one-sided t-test). Bar heights in plots indicate the mean values of n = 3 independent biological replicates. The gray line marks a ratio of 1, equivalent to the mean control fluorescence. Individual data points are indicated by black points on each plot. Error bars indicate one standard error above and below the mean values. P-values of a two-sample one-sided t-test testing if the mean of the samples are greater than the means of the controls are shown on panels a and b.
    Figure Legend Snippet: a) Fluorescence assay comparing mesobilirubin reduction activity of C. difficile CD3, C. symbiosum WAL-14163, and R. gnavus to an abiotic media controls with mesobilirubin added. b) Fluorescence assay comparing mesobilirubin reduction activity of heterologously expressed C. symbiosum bilRS in E. coli to the vector control (two-sample one-sided t-test). Bar heights in plots indicate the mean values of n = 3 independent biological replicates. The gray line marks a ratio of 1, equivalent to the mean control fluorescence. Individual data points are indicated by black points on each plot. Error bars indicate one standard error above and below the mean values. P-values of a two-sample one-sided t-test testing if the mean of the samples are greater than the means of the controls are shown on panels a and b.

    Techniques Used: Fluorescence, Activity Assay, Plasmid Preparation

    Extraction ion chromatograms for the detection of Mesobilirubin (m/z 589.3 →m/z 301.2) Urobilin (m/z 591.4→ m/z 343.2) and Stercobilin (m/z 595.4→m/z 345.2). a) Mesobilirubin standard. b) Urobilin standard. c) Stercobilin standard. d-f) E. coli transformed with pCW_gna_bilRS incubated with bilirubin (d), mesobilirubin (e) or nothing (f). g-h) C. difficile incubated with mesobilirubin (g) or nothing (h). i-j) C. symbiosum incubated with mesobilirubin (i) or nothing (J). k-l) E. coli transformed with pCW_sym_bilRS incubated with mesobilirubin (k) or nothing (l). m-n) C. symbiosum incubated with bilirubin (m) and incubated with bilirubin and co-injected with the urobilin standard (n). o-p) E. coli strains transformed with empty pCW vector incubated with bilirubin (o) and mesobilirubin (p).
    Figure Legend Snippet: Extraction ion chromatograms for the detection of Mesobilirubin (m/z 589.3 →m/z 301.2) Urobilin (m/z 591.4→ m/z 343.2) and Stercobilin (m/z 595.4→m/z 345.2). a) Mesobilirubin standard. b) Urobilin standard. c) Stercobilin standard. d-f) E. coli transformed with pCW_gna_bilRS incubated with bilirubin (d), mesobilirubin (e) or nothing (f). g-h) C. difficile incubated with mesobilirubin (g) or nothing (h). i-j) C. symbiosum incubated with mesobilirubin (i) or nothing (J). k-l) E. coli transformed with pCW_sym_bilRS incubated with mesobilirubin (k) or nothing (l). m-n) C. symbiosum incubated with bilirubin (m) and incubated with bilirubin and co-injected with the urobilin standard (n). o-p) E. coli strains transformed with empty pCW vector incubated with bilirubin (o) and mesobilirubin (p).

    Techniques Used: Extraction, Transformation Assay, Incubation, Injection, Plasmid Preparation

    a , Gene tree constructed from putative bilR sequences and related reductases from the Old Yellow Enzyme family. Experimentally confirmed bilirubin reducers are labelled in the tree. Clade 1 indicates the overall bilR clade, and clade 2 indicates the short bilR clade. The same tree with all bootstrap values and species labels is included in Extended Data Fig. . b , AlphaFold-predicted structure of the Ruminococcus gnavus BilR with sequences coloured to show their degree of conservation within clade 1 based on a ConSurf conservation analysis. A docked bilirubin (green) and FMN (orange) molecule are shown on the structure, and the insets show the potential site of interaction between the R167 residue and the putative enzyme substrates. The HGDR residues are coloured purple in the insets. c , Diagram showing the conservation of positions within the clade 1 BilR sequences. The conserved HGDR motif positions are coloured. d , Fluorescence assay results comparing the bilirubin reductase activities in E. coli 10-beta transformed with the vector control, Ruminococcus gnavus bilR and Ruminococcus gnavus bilR with the Aspartic Acid, Arginine (DR) residues at position 166–167 mutated to Alanine, Alanine (AA). The bar heights are based on the mean of n = 6 independent biological replicates used to generate the plots and statistics in d . Error bars indicate 1 s.e. above and below the mean values. P values of a two-sample one-sided t -test to determine whether the mean of the native Ruminococcus gnavus bilR was higher than the mean of the vector control or mutant are provided. Individual data points are indicated by black points on each plot.
    Figure Legend Snippet: a , Gene tree constructed from putative bilR sequences and related reductases from the Old Yellow Enzyme family. Experimentally confirmed bilirubin reducers are labelled in the tree. Clade 1 indicates the overall bilR clade, and clade 2 indicates the short bilR clade. The same tree with all bootstrap values and species labels is included in Extended Data Fig. . b , AlphaFold-predicted structure of the Ruminococcus gnavus BilR with sequences coloured to show their degree of conservation within clade 1 based on a ConSurf conservation analysis. A docked bilirubin (green) and FMN (orange) molecule are shown on the structure, and the insets show the potential site of interaction between the R167 residue and the putative enzyme substrates. The HGDR residues are coloured purple in the insets. c , Diagram showing the conservation of positions within the clade 1 BilR sequences. The conserved HGDR motif positions are coloured. d , Fluorescence assay results comparing the bilirubin reductase activities in E. coli 10-beta transformed with the vector control, Ruminococcus gnavus bilR and Ruminococcus gnavus bilR with the Aspartic Acid, Arginine (DR) residues at position 166–167 mutated to Alanine, Alanine (AA). The bar heights are based on the mean of n = 6 independent biological replicates used to generate the plots and statistics in d . Error bars indicate 1 s.e. above and below the mean values. P values of a two-sample one-sided t -test to determine whether the mean of the native Ruminococcus gnavus bilR was higher than the mean of the vector control or mutant are provided. Individual data points are indicated by black points on each plot.

    Techniques Used: Construct, Residue, Fluorescence, Transformation Assay, Plasmid Preparation, Mutagenesis

    t7 express lysy iq escherichia coli cells  (New England Biolabs)


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