ptcv-lac Search Results


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  • 99
    New England Biolabs bamhi
    Bamhi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 11236 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC cell line a549
    Cell Line A549, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega β glo assay system
    β Glo Assay System, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tecan Systems infinite m200 spectroluminometer
    Infinite M200 Spectroluminometer, supplied by Tecan Systems, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs ecori
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    92
    Thermo Fisher pcr xl topo
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    Thermo Fisher pcr blunt ii topo vector
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    ATCC chromosomal dna
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Chromosomal Dna, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 1537 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs coli t7 express lysy
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Coli T7 Express Lysy, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 2 nitrophenyl β d galactopyranoside
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    2 Nitrophenyl β D Galactopyranoside, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 181 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega beta glo assay system
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Beta Glo Assay System, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 647 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco brain heart infusion broth bhi
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Brain Heart Infusion Broth Bhi, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 143 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs ngomiv
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Ngomiv, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs eagi
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Eagi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 419 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MACHEREY NAGEL nucleospin pcr cleanup kit macherey nagel
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Nucleospin Pcr Cleanup Kit Macherey Nagel, supplied by MACHEREY NAGEL, used in various techniques. Bioz Stars score: 90/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Boehringer Mannheim primer extension
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Primer Extension, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs sticky end instant ligase
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Sticky End Instant Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Difco luria bertani broth
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Luria Bertani Broth, supplied by Difco, used in various techniques. Bioz Stars score: 93/100, based on 976 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Pel-Freez pooled rabbit serum
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Pooled Rabbit Serum, supplied by Pel-Freez, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega wizard pcr preps
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Wizard Pcr Preps, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 761 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad mueller hinton broth
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Mueller Hinton Broth, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore azocoll
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Azocoll, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim avian myeloblastosis virus reverse transcriptase
    Binding of CitO to the <t>citHO</t> - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp <t>DNA</t> probe (IR amplicon) used in the
    Avian Myeloblastosis Virus Reverse Transcriptase, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 339 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore serfa tric
    Shifts in malonyl-CoA distribution during anti-FASII adaptation. A. The diagram illustrates the two known malonyl-CoA interactants, FapR and FabD. Malonyl-CoA binds FapR (left), leading to FapR derepression. Malonyl-CoA is also a FabD substrate (right). B. Assessment of FapR-bound malonyl-CoA. FASII-antibiotic-adapted S. aureus display altered malonyl-CoA distribution. Newman expressing FapR-Trap was grown in non-selective <t>(SerFA)</t> and anti-FASII <t>(SerFA-Tric)</t> media. β-gal activities were measured in SerFA at A 600 = ∼2.0 (exponential growth: Exp, white), and in SerFA-Tric (dark grey) during latency after 3 h growth A 600 = ∼0.3 (Latency) and upon adaptive exponential outgrowth at 17 h, A 600 = ∼2 (Adapt-Exp). C. Assessment of malonyl-CoA pools. S. aureus Newman cultures were grown as in “ B ”. Left panel: Sandwich ELISA assay (green) was used to measure total malonyl-CoA (Materials and Methods). Right panel: P accBC -lacZ reporter activity (yellow). D. Assessment of FapR-bound malonyl-CoA in a fabD mutant using FapR-Trap. β-gal assays of Newman and fabD mutant (CondT R -17, a point mutant 4 ) carrying FapR-Trap were performed in SerFA after 3 h growth with A 600 = ∼1.0 (Exp; light grey). For each panel are shown the means and standard errors of three independent experiments.
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    Stratagene scs110
    Shifts in malonyl-CoA distribution during anti-FASII adaptation. A. The diagram illustrates the two known malonyl-CoA interactants, FapR and FabD. Malonyl-CoA binds FapR (left), leading to FapR derepression. Malonyl-CoA is also a FabD substrate (right). B. Assessment of FapR-bound malonyl-CoA. FASII-antibiotic-adapted S. aureus display altered malonyl-CoA distribution. Newman expressing FapR-Trap was grown in non-selective <t>(SerFA)</t> and anti-FASII <t>(SerFA-Tric)</t> media. β-gal activities were measured in SerFA at A 600 = ∼2.0 (exponential growth: Exp, white), and in SerFA-Tric (dark grey) during latency after 3 h growth A 600 = ∼0.3 (Latency) and upon adaptive exponential outgrowth at 17 h, A 600 = ∼2 (Adapt-Exp). C. Assessment of malonyl-CoA pools. S. aureus Newman cultures were grown as in “ B ”. Left panel: Sandwich ELISA assay (green) was used to measure total malonyl-CoA (Materials and Methods). Right panel: P accBC -lacZ reporter activity (yellow). D. Assessment of FapR-bound malonyl-CoA in a fabD mutant using FapR-Trap. β-gal assays of Newman and fabD mutant (CondT R -17, a point mutant 4 ) carrying FapR-Trap were performed in SerFA after 3 h growth with A 600 = ∼1.0 (Exp; light grey). For each panel are shown the means and standard errors of three independent experiments.
    Scs110, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Binding of CitO to the citHO - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp DNA probe (IR amplicon) used in the

    Journal:

    Article Title: Transcriptional Regulation of the Citrate Gene Cluster of Enterococcus faecalis Involves the GntR Family Transcriptional Activator CitO ▿ Involves the GntR Family Transcriptional Activator CitO ▿ †

    doi: 10.1128/JB.01704-07

    Figure Lengend Snippet: Binding of CitO to the citHO - oadHDB - citCDEFX - oadA - citMG IR. (A) Schematic representation of the cit operon IR. P citHO and P citCL promoters are indicated. The thick line on the top indicates the location of the 427-bp DNA probe (IR amplicon) used in the

    Article Snippet: To construct the pTCV- lac -derivative plasmids, DNA fragments from the promoter regions of the citHO and oadHDB-citCDEFX-oadA-citMG operons were obtained by PCR using chromosomal DNA of E. faecalis ATCC 29212 as the template and cloned into the pCR-Blunt II-TOPO vector (Invitrogen). pTOPO-derivative plasmids were digested with EcoRI, and each released fragment was ligated into the corresponding site of pTCV- lac vector.

    Techniques: Binding Assay, Amplification

    (A) Effect of diverse citrate analogs on DNase I protection of the cit promoters by CitO. The oadHDB - citCDEFX - oadA - citMG coding strand was employed as target DNA. The CitO protein was used at a concentration of 1.5 μM. The different citrate analogs

    Journal:

    Article Title: Transcriptional Regulation of the Citrate Gene Cluster of Enterococcus faecalis Involves the GntR Family Transcriptional Activator CitO ▿ Involves the GntR Family Transcriptional Activator CitO ▿ †

    doi: 10.1128/JB.01704-07

    Figure Lengend Snippet: (A) Effect of diverse citrate analogs on DNase I protection of the cit promoters by CitO. The oadHDB - citCDEFX - oadA - citMG coding strand was employed as target DNA. The CitO protein was used at a concentration of 1.5 μM. The different citrate analogs

    Article Snippet: To construct the pTCV- lac -derivative plasmids, DNA fragments from the promoter regions of the citHO and oadHDB-citCDEFX-oadA-citMG operons were obtained by PCR using chromosomal DNA of E. faecalis ATCC 29212 as the template and cloned into the pCR-Blunt II-TOPO vector (Invitrogen). pTOPO-derivative plasmids were digested with EcoRI, and each released fragment was ligated into the corresponding site of pTCV- lac vector.

    Techniques: Concentration Assay

    DNase I protection experiments of the operons promoters by CitO. A 427-bp DNA fragment including the citHO - oadHDB - citCDEFX - oadA - citMG operon promoter regions was end labeled with [γ- 32 P]ATP and used as target DNA in the assays. Each strand was

    Journal:

    Article Title: Transcriptional Regulation of the Citrate Gene Cluster of Enterococcus faecalis Involves the GntR Family Transcriptional Activator CitO ▿ Involves the GntR Family Transcriptional Activator CitO ▿ †

    doi: 10.1128/JB.01704-07

    Figure Lengend Snippet: DNase I protection experiments of the operons promoters by CitO. A 427-bp DNA fragment including the citHO - oadHDB - citCDEFX - oadA - citMG operon promoter regions was end labeled with [γ- 32 P]ATP and used as target DNA in the assays. Each strand was

    Article Snippet: To construct the pTCV- lac -derivative plasmids, DNA fragments from the promoter regions of the citHO and oadHDB-citCDEFX-oadA-citMG operons were obtained by PCR using chromosomal DNA of E. faecalis ATCC 29212 as the template and cloned into the pCR-Blunt II-TOPO vector (Invitrogen). pTOPO-derivative plasmids were digested with EcoRI, and each released fragment was ligated into the corresponding site of pTCV- lac vector.

    Techniques: Labeling

    Shifts in malonyl-CoA distribution during anti-FASII adaptation. A. The diagram illustrates the two known malonyl-CoA interactants, FapR and FabD. Malonyl-CoA binds FapR (left), leading to FapR derepression. Malonyl-CoA is also a FabD substrate (right). B. Assessment of FapR-bound malonyl-CoA. FASII-antibiotic-adapted S. aureus display altered malonyl-CoA distribution. Newman expressing FapR-Trap was grown in non-selective (SerFA) and anti-FASII (SerFA-Tric) media. β-gal activities were measured in SerFA at A 600 = ∼2.0 (exponential growth: Exp, white), and in SerFA-Tric (dark grey) during latency after 3 h growth A 600 = ∼0.3 (Latency) and upon adaptive exponential outgrowth at 17 h, A 600 = ∼2 (Adapt-Exp). C. Assessment of malonyl-CoA pools. S. aureus Newman cultures were grown as in “ B ”. Left panel: Sandwich ELISA assay (green) was used to measure total malonyl-CoA (Materials and Methods). Right panel: P accBC -lacZ reporter activity (yellow). D. Assessment of FapR-bound malonyl-CoA in a fabD mutant using FapR-Trap. β-gal assays of Newman and fabD mutant (CondT R -17, a point mutant 4 ) carrying FapR-Trap were performed in SerFA after 3 h growth with A 600 = ∼1.0 (Exp; light grey). For each panel are shown the means and standard errors of three independent experiments.

    Journal: bioRxiv

    Article Title: (p)ppGpp and malonyl-CoA set the pace for Staphylococcus aureus adaptation to FASII antibiotics and provide a basis for bi-therapy inhibition

    doi: 10.1101/2020.03.26.007567

    Figure Lengend Snippet: Shifts in malonyl-CoA distribution during anti-FASII adaptation. A. The diagram illustrates the two known malonyl-CoA interactants, FapR and FabD. Malonyl-CoA binds FapR (left), leading to FapR derepression. Malonyl-CoA is also a FabD substrate (right). B. Assessment of FapR-bound malonyl-CoA. FASII-antibiotic-adapted S. aureus display altered malonyl-CoA distribution. Newman expressing FapR-Trap was grown in non-selective (SerFA) and anti-FASII (SerFA-Tric) media. β-gal activities were measured in SerFA at A 600 = ∼2.0 (exponential growth: Exp, white), and in SerFA-Tric (dark grey) during latency after 3 h growth A 600 = ∼0.3 (Latency) and upon adaptive exponential outgrowth at 17 h, A 600 = ∼2 (Adapt-Exp). C. Assessment of malonyl-CoA pools. S. aureus Newman cultures were grown as in “ B ”. Left panel: Sandwich ELISA assay (green) was used to measure total malonyl-CoA (Materials and Methods). Right panel: P accBC -lacZ reporter activity (yellow). D. Assessment of FapR-bound malonyl-CoA in a fabD mutant using FapR-Trap. β-gal assays of Newman and fabD mutant (CondT R -17, a point mutant 4 ) carrying FapR-Trap were performed in SerFA after 3 h growth with A 600 = ∼1.0 (Exp; light grey). For each panel are shown the means and standard errors of three independent experiments.

    Article Snippet: Ser-FA (BHI containing eFA+10% newborn calf serum; Sigma) and SerFA-Tric (SerFA+Triclosan 0.5 µg/ml) were the modified media used as indicated.

    Techniques: Expressing, Sandwich ELISA, Activity Assay, Mutagenesis

    Expression of PlsX and PlsC in FASII-antibiotic-induced latency and adaptation. A. Sequence alignment of FapR binding sites in S. aureus N315 using the RegPrecise database 13 , 34 . Positions correspond to the predicted start codon of the first Orf in each operon. Consensus nucleotides in the 17 bp FapR binding sequence are shaded. In the consensus sequence below the alignment: N, any nucleotide; W, A or T; Y, T or C. Putative -10 RNA polymerase binding sites are underlined (from 13 ). The PlsC ATG and FapR TTG start sites are in italics. B, C. Shifts in plsX and plsC expression during growth in non-selective (SerFA) and anti-FASII (SerFA-Tric) media. β-gal activities of S. aureus Newman expressing P fapR plsX -lacZ and P plsC -lacZ are shown for samples processed in SerFA at OD 600 = ∼2.0 (exponential phase, Exp), and in SerFA-Tric after 3h in latency (OD 600 = ∼0.3) and upon adaptive exponential outgrowth at 17 h (OD 600 = ∼2.0; Adapt-Exp). Non-selective growth, white; selective growth, dark grey. For each panel are shown the means and standard errors of three independent experiments.

    Journal: bioRxiv

    Article Title: (p)ppGpp and malonyl-CoA set the pace for Staphylococcus aureus adaptation to FASII antibiotics and provide a basis for bi-therapy inhibition

    doi: 10.1101/2020.03.26.007567

    Figure Lengend Snippet: Expression of PlsX and PlsC in FASII-antibiotic-induced latency and adaptation. A. Sequence alignment of FapR binding sites in S. aureus N315 using the RegPrecise database 13 , 34 . Positions correspond to the predicted start codon of the first Orf in each operon. Consensus nucleotides in the 17 bp FapR binding sequence are shaded. In the consensus sequence below the alignment: N, any nucleotide; W, A or T; Y, T or C. Putative -10 RNA polymerase binding sites are underlined (from 13 ). The PlsC ATG and FapR TTG start sites are in italics. B, C. Shifts in plsX and plsC expression during growth in non-selective (SerFA) and anti-FASII (SerFA-Tric) media. β-gal activities of S. aureus Newman expressing P fapR plsX -lacZ and P plsC -lacZ are shown for samples processed in SerFA at OD 600 = ∼2.0 (exponential phase, Exp), and in SerFA-Tric after 3h in latency (OD 600 = ∼0.3) and upon adaptive exponential outgrowth at 17 h (OD 600 = ∼2.0; Adapt-Exp). Non-selective growth, white; selective growth, dark grey. For each panel are shown the means and standard errors of three independent experiments.

    Article Snippet: Ser-FA (BHI containing eFA+10% newborn calf serum; Sigma) and SerFA-Tric (SerFA+Triclosan 0.5 µg/ml) were the modified media used as indicated.

    Techniques: Expressing, Sequencing, Binding Assay

    S. aureus bypasses FASII inhibition by exogenous fatty acid (eFA) incorporation in membrane phospholipids. A. Model for anti-FASII adaptation. FASII and FASII bypass are schematized as characterized; functions whose expression is controlled by FapR repressor are underlined 16 - 19 . eFA phosphorylation by Fak (fatty acid kinase) 20 provides an intermediate that may either be incorporated in position 1 of the glycerol-3-phosphate backbone via PlsY, or act as a substrate for PlsX to then be incorporated in position 2 via PlsC. In the absence of serum, FabD (malonyl-CoA:ACP transacylase) mutations promote anti-FASII adaptation 16 . In contrast, serum favors FASII antibiotic adaptation without FASII mutations 21 . B. Fatty acid profiles of S. aureus Newman. Left, BHI grown cells; right, cells grown overnight in SerFA-Tric. Cultures started at A 600 = 0.01 were harvested at A 600 = 1. Arrow indicates anteiso 15 ( ai 15), the major fatty acid synthesized by S. aureus . eFA: 1, C14:0; 2, C16:0; 3, C18:1. Profiles are representative of three independent experiments. FA, fatty acids; FA-ACP, fatty acyl-ACP; FA-PO 4 , acyl-phosphate; grey, inhibited pathway. FabD * , mutated or inhibited enzymes.

    Journal: bioRxiv

    Article Title: (p)ppGpp and malonyl-CoA set the pace for Staphylococcus aureus adaptation to FASII antibiotics and provide a basis for bi-therapy inhibition

    doi: 10.1101/2020.03.26.007567

    Figure Lengend Snippet: S. aureus bypasses FASII inhibition by exogenous fatty acid (eFA) incorporation in membrane phospholipids. A. Model for anti-FASII adaptation. FASII and FASII bypass are schematized as characterized; functions whose expression is controlled by FapR repressor are underlined 16 - 19 . eFA phosphorylation by Fak (fatty acid kinase) 20 provides an intermediate that may either be incorporated in position 1 of the glycerol-3-phosphate backbone via PlsY, or act as a substrate for PlsX to then be incorporated in position 2 via PlsC. In the absence of serum, FabD (malonyl-CoA:ACP transacylase) mutations promote anti-FASII adaptation 16 . In contrast, serum favors FASII antibiotic adaptation without FASII mutations 21 . B. Fatty acid profiles of S. aureus Newman. Left, BHI grown cells; right, cells grown overnight in SerFA-Tric. Cultures started at A 600 = 0.01 were harvested at A 600 = 1. Arrow indicates anteiso 15 ( ai 15), the major fatty acid synthesized by S. aureus . eFA: 1, C14:0; 2, C16:0; 3, C18:1. Profiles are representative of three independent experiments. FA, fatty acids; FA-ACP, fatty acyl-ACP; FA-PO 4 , acyl-phosphate; grey, inhibited pathway. FabD * , mutated or inhibited enzymes.

    Article Snippet: Ser-FA (BHI containing eFA+10% newborn calf serum; Sigma) and SerFA-Tric (SerFA+Triclosan 0.5 µg/ml) were the modified media used as indicated.

    Techniques: Inhibition, Expressing, Synthesized

    Subinhibitory mupirocin addition inhibits S. aureus eFA incorporation in membrane phospholipids and synergizes with anti-FASII to inhibit adaptation. A , Left, Fatty acid profiles of S. aureus Newman grown in SerFA and SerFA+mupirocin (Mupi). Samples were processed after 3 h growth. Black arrow, position of the main endogenous fatty acid ai 15:0. eFA are; 1- C14:0; 2- C16:0; 3- C18:1; 4- C20:1 (elongation of C18:1). Dashed arrow, elongation of C18:1 (n+2). Right, percent eFA of Newman grown in SerFA without and with Mupi, derived from integration of fatty acid profiles. B. Left, S. aureus Newman cultures were grown in SerFA and SerFA+Mupi, or right, in SerFA-Tric and SerFA-Tric+Mupi. Growth was monitored by A 600 . Growth curves at right are shown starting at 10 h. Mupi was used at 0.05 µg/ml. Results are representative of three independent experiments.

    Journal: bioRxiv

    Article Title: (p)ppGpp and malonyl-CoA set the pace for Staphylococcus aureus adaptation to FASII antibiotics and provide a basis for bi-therapy inhibition

    doi: 10.1101/2020.03.26.007567

    Figure Lengend Snippet: Subinhibitory mupirocin addition inhibits S. aureus eFA incorporation in membrane phospholipids and synergizes with anti-FASII to inhibit adaptation. A , Left, Fatty acid profiles of S. aureus Newman grown in SerFA and SerFA+mupirocin (Mupi). Samples were processed after 3 h growth. Black arrow, position of the main endogenous fatty acid ai 15:0. eFA are; 1- C14:0; 2- C16:0; 3- C18:1; 4- C20:1 (elongation of C18:1). Dashed arrow, elongation of C18:1 (n+2). Right, percent eFA of Newman grown in SerFA without and with Mupi, derived from integration of fatty acid profiles. B. Left, S. aureus Newman cultures were grown in SerFA and SerFA+Mupi, or right, in SerFA-Tric and SerFA-Tric+Mupi. Growth was monitored by A 600 . Growth curves at right are shown starting at 10 h. Mupi was used at 0.05 µg/ml. Results are representative of three independent experiments.

    Article Snippet: Ser-FA (BHI containing eFA+10% newborn calf serum; Sigma) and SerFA-Tric (SerFA+Triclosan 0.5 µg/ml) were the modified media used as indicated.

    Techniques: Derivative Assay

    Transient S. aureus stringent response induction during the latency phase prior to anti-FASII adaptation. A. Growth kinetics of S. aureus Newman in BHI (black line) and in SerFA-Tric (green) 5 . In SerFA-Tric, a 10-12 h latency period (Latency) precedes adaptive exponential outgrowth (Adapt-Exp). B. Stringent response status according to growth condition in Newman carrying P oppB -lacZ reporter (blue, left panel) and P cshA -lacZ (pink, right panel). β-gal activities of samples in SerFA (exponential growth; Exp, A 600 = ∼1.5) and SerFA-Tric (Latency, A 600 = ∼0.3) are measured after 3 h growth. β-gal activities were measured on 17 h samples in SerFA-Tric (exponential growth; Adapt-Exp, A 600 = ∼1.5). Means of three independent cultures with standard errors (SE; not visible for BHI growth) are shown for each experiment.

    Journal: bioRxiv

    Article Title: (p)ppGpp and malonyl-CoA set the pace for Staphylococcus aureus adaptation to FASII antibiotics and provide a basis for bi-therapy inhibition

    doi: 10.1101/2020.03.26.007567

    Figure Lengend Snippet: Transient S. aureus stringent response induction during the latency phase prior to anti-FASII adaptation. A. Growth kinetics of S. aureus Newman in BHI (black line) and in SerFA-Tric (green) 5 . In SerFA-Tric, a 10-12 h latency period (Latency) precedes adaptive exponential outgrowth (Adapt-Exp). B. Stringent response status according to growth condition in Newman carrying P oppB -lacZ reporter (blue, left panel) and P cshA -lacZ (pink, right panel). β-gal activities of samples in SerFA (exponential growth; Exp, A 600 = ∼1.5) and SerFA-Tric (Latency, A 600 = ∼0.3) are measured after 3 h growth. β-gal activities were measured on 17 h samples in SerFA-Tric (exponential growth; Adapt-Exp, A 600 = ∼1.5). Means of three independent cultures with standard errors (SE; not visible for BHI growth) are shown for each experiment.

    Article Snippet: Ser-FA (BHI containing eFA+10% newborn calf serum; Sigma) and SerFA-Tric (SerFA+Triclosan 0.5 µg/ml) were the modified media used as indicated.

    Techniques: