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VectorBuilder GmbH pbad-flc
Pbad Flc, supplied by VectorBuilder GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Related Articles

Plasmid Preparation:

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: Because the plasmid carrying the gene could not be transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, pBAD-FLC (Vectorbuilder), and expressed in E. coli LMG194 (pBAD TOP TA expression kit, Invitrogen, Saint-Aubin, France) ( ).

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: As the plasmid encoding the gene could not be 75 transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, 76 pBAD-FLC (Vectorbuilder) and expressed in E. coli LMG194 (pBAD TOP TA Expression Kit, Invitrogen, Saint-77 Aubin France) (15).

Transformation Assay:

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: Because the plasmid carrying the gene could not be transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, pBAD-FLC (Vectorbuilder), and expressed in E. coli LMG194 (pBAD TOP TA expression kit, Invitrogen, Saint-Aubin, France) ( ).

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: As the plasmid encoding the gene could not be 75 transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, 76 pBAD-FLC (Vectorbuilder) and expressed in E. coli LMG194 (pBAD TOP TA Expression Kit, Invitrogen, Saint-77 Aubin France) (15).

Clone Assay:

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: Because the plasmid carrying the gene could not be transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, pBAD-FLC (Vectorbuilder), and expressed in E. coli LMG194 (pBAD TOP TA expression kit, Invitrogen, Saint-Aubin, France) ( ).

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: As the plasmid encoding the gene could not be 75 transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, 76 pBAD-FLC (Vectorbuilder) and expressed in E. coli LMG194 (pBAD TOP TA Expression Kit, Invitrogen, Saint-77 Aubin France) (15).

Expressing:

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: Because the plasmid carrying the gene could not be transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, pBAD-FLC (Vectorbuilder), and expressed in E. coli LMG194 (pBAD TOP TA expression kit, Invitrogen, Saint-Aubin, France) ( ).

Article Title: Novel Carbapenemases FLC-1 and IMI-2 Encoded by an Enterobacter cloacae Complex Isolated from Food Products
Article Snippet: As the plasmid encoding the gene could not be 75 transformed or conjugated into E. coli cells, the gene was cloned into an arabinose-inducible expression vector, 76 pBAD-FLC (Vectorbuilder) and expressed in E. coli LMG194 (pBAD TOP TA Expression Kit, Invitrogen, Saint-77 Aubin France) (15).



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SLA1 deletion drives FLC resistance through upregulation of CDR1. (A) Spotting assays of WT and sla1Δ mutants. Strains were spotted onto YPD agar supplemented with the FLC, KTC, AMB and the plates were incubated at 30 for 2 days. (B) RT-qPCR analysis of ERG genes, CDR1 , CDR2 , MDR1 and PMC1 in WT and sla1Δ strains ( n = 3). (C) Spotting assays of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. Indicated strains were spotted with or without FLC. (D) FLC IC₅₀ values of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. (E) Correlation analysis between SLA1 and CDR1 transcript levels in 17 clinical C. auris isolates. (F) RT-qPCR analysis of CDR1 regulators CAP1 , TAC1 , and GSK3 in WT and sla1Δ strains ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Journal: Frontiers in Microbiology

Article Title: SLA1 governs azole resistance in Candidozyma auris via mitochondrial ROS-driven efflux pump expression

doi: 10.3389/fmicb.2026.1948008

Figure Lengend Snippet: SLA1 deletion drives FLC resistance through upregulation of CDR1. (A) Spotting assays of WT and sla1Δ mutants. Strains were spotted onto YPD agar supplemented with the FLC, KTC, AMB and the plates were incubated at 30 for 2 days. (B) RT-qPCR analysis of ERG genes, CDR1 , CDR2 , MDR1 and PMC1 in WT and sla1Δ strains ( n = 3). (C) Spotting assays of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. Indicated strains were spotted with or without FLC. (D) FLC IC₅₀ values of WT, sla1Δ , cdr1Δ , and sla1Δcdr1Δ strains. (E) Correlation analysis between SLA1 and CDR1 transcript levels in 17 clinical C. auris isolates. (F) RT-qPCR analysis of CDR1 regulators CAP1 , TAC1 , and GSK3 in WT and sla1Δ strains ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Article Snippet: Drug-resistance tests were performed using 20 μg/mL FLC (MedChemExpress, cat# HY-B0101), 0.2 μg/mL ketoconazole (KTC) (MedChemExpress, cat# HY-B0105), 1.0 μg/mL Amphotericin B (AMB) (MedChemExpress, cat# HY-B0221), 10 mM NAC (Sigma, cat# 1009005), 20 μM Antimycin A (Sigma, cat# 77332), 50 μM Mito-TEMPO (MedChemExpress, cat# HY-112879), 60 μM Rotenone (MedChemExpress, cat# HY-B1756), and 1 mM H2O2 (Sigma, cat# 108600).

Techniques: Incubation, Quantitative RT-PCR, Analysis, Two Tailed Test

SLA1 deletion disrupts redox homeostasis and mitochondrial energy metabolism to regulate FLC resistance in C. auris . (A) GO analyses. GO term enrichment analyses were performed using the RNA-seq data from WT and sla1Δ strains. (B,C) Gene Set Enrichment Analysis (GSEA). Comparing the transcriptome data from the WT and sla1Δ strains. (D) Spotting assays of WT and sla1Δ strains. Strains were spotted onto YPD agar supplemented FLC, FLC + NAC, or FLC + H₂O₂ conditions. (E) Flow cytometry analysis of intracellular ROS levels in WT, and sla1Δ strains. Wild-type and sla1Δ cells were cultured YPD, loaded with MitoSOX, and then analyzed by flow cytometry. (F) Analysis of ROS levels. WT and sla1Δ strains were incubated at 30 °C for 12 h, then ROS levels were measured ( n = 4). (G) Analysis of ATP levels. WT and sla1Δ strains were incubated at 30 °C, then ATP levels were measured ( n = 3). (H) RT-qPCR analysis of genes involved in mitochondrial respiration and energy metabolism in WT and sla1Δ strains ( n = 3). (I) Analysis of ATP and ROS levels. Measurement of relative intracellular ATP and ROS content in laboratory wild-type susceptible strain and five clinical fluconazole-resistant C. auris isolates. ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Journal: Frontiers in Microbiology

Article Title: SLA1 governs azole resistance in Candidozyma auris via mitochondrial ROS-driven efflux pump expression

doi: 10.3389/fmicb.2026.1948008

Figure Lengend Snippet: SLA1 deletion disrupts redox homeostasis and mitochondrial energy metabolism to regulate FLC resistance in C. auris . (A) GO analyses. GO term enrichment analyses were performed using the RNA-seq data from WT and sla1Δ strains. (B,C) Gene Set Enrichment Analysis (GSEA). Comparing the transcriptome data from the WT and sla1Δ strains. (D) Spotting assays of WT and sla1Δ strains. Strains were spotted onto YPD agar supplemented FLC, FLC + NAC, or FLC + H₂O₂ conditions. (E) Flow cytometry analysis of intracellular ROS levels in WT, and sla1Δ strains. Wild-type and sla1Δ cells were cultured YPD, loaded with MitoSOX, and then analyzed by flow cytometry. (F) Analysis of ROS levels. WT and sla1Δ strains were incubated at 30 °C for 12 h, then ROS levels were measured ( n = 4). (G) Analysis of ATP levels. WT and sla1Δ strains were incubated at 30 °C, then ATP levels were measured ( n = 3). (H) RT-qPCR analysis of genes involved in mitochondrial respiration and energy metabolism in WT and sla1Δ strains ( n = 3). (I) Analysis of ATP and ROS levels. Measurement of relative intracellular ATP and ROS content in laboratory wild-type susceptible strain and five clinical fluconazole-resistant C. auris isolates. ( n = 3). Two-tailed unpaired t -tests were used for statistical analyses; data are expressed as the mean ± SD.

Article Snippet: Drug-resistance tests were performed using 20 μg/mL FLC (MedChemExpress, cat# HY-B0101), 0.2 μg/mL ketoconazole (KTC) (MedChemExpress, cat# HY-B0105), 1.0 μg/mL Amphotericin B (AMB) (MedChemExpress, cat# HY-B0221), 10 mM NAC (Sigma, cat# 1009005), 20 μM Antimycin A (Sigma, cat# 77332), 50 μM Mito-TEMPO (MedChemExpress, cat# HY-112879), 60 μM Rotenone (MedChemExpress, cat# HY-B1756), and 1 mM H2O2 (Sigma, cat# 108600).

Techniques: RNA Sequencing, Analysis, Flow Cytometry, Cell Culture, Incubation, Quantitative RT-PCR, Two Tailed Test

Box plots illustrating (a) the number of CSF-specific oligoclonal bands (OCB) of OCB positive persons with MS and CIS, (b) intrathecal produced amount of IgG of persons with MS and CIS with detectable IgG-synthesis, according to Reiber’s diagram, and (c) KFLC index of all persons with MS and CIS with KFLC index >6.1. Panel (d) shows OCB count cross across different clinical presentation types. Boxes represent interquartile range with horizontal line within the box representing the median. Lower and upper whiskers represent min. and max. values. MS, multiple sclerosis; CIS, clinically isolated syndrome; OCB, oligoclonal bands; CSF, cerebrospinal fluid; IgG, immunoglobulin G; KFLC, kappa free light chains.

Journal: Frontiers in Neurology

Article Title: Assessing the value of oligoclonal band count in multiple sclerosis: insights from a large cohort analysis

doi: 10.3389/fneur.2026.1719470

Figure Lengend Snippet: Box plots illustrating (a) the number of CSF-specific oligoclonal bands (OCB) of OCB positive persons with MS and CIS, (b) intrathecal produced amount of IgG of persons with MS and CIS with detectable IgG-synthesis, according to Reiber’s diagram, and (c) KFLC index of all persons with MS and CIS with KFLC index >6.1. Panel (d) shows OCB count cross across different clinical presentation types. Boxes represent interquartile range with horizontal line within the box representing the median. Lower and upper whiskers represent min. and max. values. MS, multiple sclerosis; CIS, clinically isolated syndrome; OCB, oligoclonal bands; CSF, cerebrospinal fluid; IgG, immunoglobulin G; KFLC, kappa free light chains.

Article Snippet: Serum and CSF concentrations of KFLC were measured using nephelometry with the N Latex FLC kappa kit (Siemens Healthcare Diagnostics Products GmbH), following the manufacturer’s instructions on an Atellica Neph 630 System (Siemens Healthcare Diagnostics Products GmbH).

Techniques: Produced, Isolation

Receiver operating characteristic (ROC) curves for the differentiation of MS and CIS ( n = 269). The plot illustrates the diagnostic accuracy of OCB count, KFLC index, intrathecal IgG synthesis according to Reiber, and the combination of OCB count and KFLC index. The area under the curve (AUC) with corresponding 95% confidence interval (CI) for each methodology is indicated in the legend. The distinct inflection point in the intrathecal IgG curve reflects the high proportion of individuals with a calculated IgG synthesis of 0.0 mg/L, where the methodology reaches its limit of continuous discrimination. MS, multiple sclerosis; CIS, clinically isolated syndrome; OCB, oligoclonal bands; KFLC, kappa free light chain; IgG, immunoglobulin G.

Journal: Frontiers in Neurology

Article Title: Assessing the value of oligoclonal band count in multiple sclerosis: insights from a large cohort analysis

doi: 10.3389/fneur.2026.1719470

Figure Lengend Snippet: Receiver operating characteristic (ROC) curves for the differentiation of MS and CIS ( n = 269). The plot illustrates the diagnostic accuracy of OCB count, KFLC index, intrathecal IgG synthesis according to Reiber, and the combination of OCB count and KFLC index. The area under the curve (AUC) with corresponding 95% confidence interval (CI) for each methodology is indicated in the legend. The distinct inflection point in the intrathecal IgG curve reflects the high proportion of individuals with a calculated IgG synthesis of 0.0 mg/L, where the methodology reaches its limit of continuous discrimination. MS, multiple sclerosis; CIS, clinically isolated syndrome; OCB, oligoclonal bands; KFLC, kappa free light chain; IgG, immunoglobulin G.

Article Snippet: Serum and CSF concentrations of KFLC were measured using nephelometry with the N Latex FLC kappa kit (Siemens Healthcare Diagnostics Products GmbH), following the manufacturer’s instructions on an Atellica Neph 630 System (Siemens Healthcare Diagnostics Products GmbH).

Techniques: Diagnostic Assay, Isolation

Scatter plots showing the positive correlations between the number of oligoclonal bands (OCB) in CSF with (a) intrathecal IgG levels (rho = 0.6963, p < 0.0001) and (b) KFLC index (rho = 0.7099, p < 0.0001). MS, multiple sclerosis; OCB, oligoclonal bands; CSF, cerebrospinal fluid; KFLC, kappa free light chains; EDSS, IgG, immunoglobulin G.

Journal: Frontiers in Neurology

Article Title: Assessing the value of oligoclonal band count in multiple sclerosis: insights from a large cohort analysis

doi: 10.3389/fneur.2026.1719470

Figure Lengend Snippet: Scatter plots showing the positive correlations between the number of oligoclonal bands (OCB) in CSF with (a) intrathecal IgG levels (rho = 0.6963, p < 0.0001) and (b) KFLC index (rho = 0.7099, p < 0.0001). MS, multiple sclerosis; OCB, oligoclonal bands; CSF, cerebrospinal fluid; KFLC, kappa free light chains; EDSS, IgG, immunoglobulin G.

Article Snippet: Serum and CSF concentrations of KFLC were measured using nephelometry with the N Latex FLC kappa kit (Siemens Healthcare Diagnostics Products GmbH), following the manufacturer’s instructions on an Atellica Neph 630 System (Siemens Healthcare Diagnostics Products GmbH).

Techniques: