β lactamase type iv  (Millipore)

 
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    Name:
    beta Lactamase
    Description:
    β Lactamase produced by bacteria is closely related to the penicillin binding proteins β Lactamase hydrolyzes β lactum antibiotics and is the prime cause of resistance development by bacteria There are four subclasses of β Lactamases Classes A C and D form an acyl enzyme via active site serine residue Class B β lactamases are metalloenzymes with zinc ion at their active site for β lactam hydrolysis Mutations in the β lactamases has resulted in the generation of extended spectrum β lactamases ESBLs As close to 900 types of β Lactamases are produced by microbes
    Catalog Number:
    L6170
    Price:
    None
    Applications:
    β--lactamase is used to inactivate β-lactam antibiotics by breaking open the β-lactam ring. β--lactamase is used to study antibiotic resistance and resistance suppression. Product L6170 is recombinantly produced based on the sequence of the enzyme from Pseudomonas aeruginosa, and is expressed in E. coli.
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    Structured Review

    Millipore β lactamase type iv
    The inhibitory activity of SSE against <t>β</t> -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p
    β Lactamase produced by bacteria is closely related to the penicillin binding proteins β Lactamase hydrolyzes β lactum antibiotics and is the prime cause of resistance development by bacteria There are four subclasses of β Lactamases Classes A C and D form an acyl enzyme via active site serine residue Class B β lactamases are metalloenzymes with zinc ion at their active site for β lactam hydrolysis Mutations in the β lactamases has resulted in the generation of extended spectrum β lactamases ESBLs As close to 900 types of β Lactamases are produced by microbes
    https://www.bioz.com/result/β lactamase type iv/product/Millipore
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    β lactamase type iv - by Bioz Stars, 2021-07
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    Images

    1) Product Images from "The Synergy and Mode of Action of Cyperus rotundus L. Extract Plus Ampicillin against Ampicillin-Resistant Staphylococcus aureus"

    Article Title: The Synergy and Mode of Action of Cyperus rotundus L. Extract Plus Ampicillin against Ampicillin-Resistant Staphylococcus aureus

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2018/3438453

    The inhibitory activity of SSE against β -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p
    Figure Legend Snippet: The inhibitory activity of SSE against β -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p

    Techniques Used: Activity Assay

    2) Product Images from "Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus"

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus

    Journal: Antimicrobial Agents and Chemotherapy

    doi:

    Agarose gel (a) and Southern hybridization with a blaZ probe (b) of Eco RI-restricted plasmid DNA from phage group II, β-lactamase-producing isolates of S. aureus ).
    Figure Legend Snippet: Agarose gel (a) and Southern hybridization with a blaZ probe (b) of Eco RI-restricted plasmid DNA from phage group II, β-lactamase-producing isolates of S. aureus ).

    Techniques Used: Agarose Gel Electrophoresis, Hybridization, Plasmid Preparation

    Southern hybridization of chromosomal DNA from phage group II, type B β-lactamase-producing isolates of S. aureus using a blaZ probe. Lanes 1 to 9, Eco RI-restricted DNA; lanes 10 to 18, Hin ).
    Figure Legend Snippet: Southern hybridization of chromosomal DNA from phage group II, type B β-lactamase-producing isolates of S. aureus using a blaZ probe. Lanes 1 to 9, Eco RI-restricted DNA; lanes 10 to 18, Hin ).

    Techniques Used: Hybridization

    Comparison of the nucleotide sequence of the gene encoding type B β-lactamase from strain 22260 to other S. aureus ). Differences in nucleotide sequences are shown; −, no change from the blaZ ). L1 to L24 refer to the leader peptide. Whereas the sequences of the type A, C, and D β-lactamases have been determined through the carboxy-terminal residue at Ambler position 290, reliable sequence data for the type B enzyme were obtained through residue 253.
    Figure Legend Snippet: Comparison of the nucleotide sequence of the gene encoding type B β-lactamase from strain 22260 to other S. aureus ). Differences in nucleotide sequences are shown; −, no change from the blaZ ). L1 to L24 refer to the leader peptide. Whereas the sequences of the type A, C, and D β-lactamases have been determined through the carboxy-terminal residue at Ambler position 290, reliable sequence data for the type B enzyme were obtained through residue 253.

    Techniques Used: Sequencing

    3) Product Images from "The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone"

    Article Title: The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.186.11.3621-3630.2004

    HopPtoV is secreted in culture via the DC3000 TTSS in an ShcV-dependent manner. (A) Cultures of wild-type (WT) P. syringae pv. tomato DC3000 and a DC3000 TTSS-defective mutant ( hrcC ) were grown in hrp -inducing medium and separated into cell-bound (C) and supernatant (S) fractions. The cell-bound and supernatant fractions were concentrated 13.3-fold and 133-fold, respectively, relative to the initial culture volumes. The samples were separated by SDS-PAGE and immunoblotted. HopPtoV and β-lactamase, which was used as a lysis control, were detected with anti-HA and anti-β-lactamase antibodies, respectively. Wild-type DC3000 strains containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) secreted HopPtoV-HA into the supernatant fraction. However, much more HopPtoV-HA was found in the supernatant fractions from DC3000(pLN517), which overexpressed ShcV. (B) Cultures of DC3000 shcV (UNL120) and hopPtoV (UNL125) insertion mutants containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) were grown in hrp -inducing medium, separated into cell-bound (C) and supernatant (S) fractions, and separated by SDS-PAGE. Immunoblotted proteins were detected with anti-HA or anti-NPTII antibodies. Plasmid-encoded NPTII should remain cell bound and was used as a lysis control. The shcV mutant UNL120 was unable to secrete HopPtoV-HA unless shcV was also provided in trans .
    Figure Legend Snippet: HopPtoV is secreted in culture via the DC3000 TTSS in an ShcV-dependent manner. (A) Cultures of wild-type (WT) P. syringae pv. tomato DC3000 and a DC3000 TTSS-defective mutant ( hrcC ) were grown in hrp -inducing medium and separated into cell-bound (C) and supernatant (S) fractions. The cell-bound and supernatant fractions were concentrated 13.3-fold and 133-fold, respectively, relative to the initial culture volumes. The samples were separated by SDS-PAGE and immunoblotted. HopPtoV and β-lactamase, which was used as a lysis control, were detected with anti-HA and anti-β-lactamase antibodies, respectively. Wild-type DC3000 strains containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) secreted HopPtoV-HA into the supernatant fraction. However, much more HopPtoV-HA was found in the supernatant fractions from DC3000(pLN517), which overexpressed ShcV. (B) Cultures of DC3000 shcV (UNL120) and hopPtoV (UNL125) insertion mutants containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) were grown in hrp -inducing medium, separated into cell-bound (C) and supernatant (S) fractions, and separated by SDS-PAGE. Immunoblotted proteins were detected with anti-HA or anti-NPTII antibodies. Plasmid-encoded NPTII should remain cell bound and was used as a lysis control. The shcV mutant UNL120 was unable to secrete HopPtoV-HA unless shcV was also provided in trans .

    Techniques Used: Mutagenesis, SDS Page, Lysis, Plasmid Preparation

    4) Product Images from "ClbP Is a Prototype of a Peptidase Subgroup Involved in Biosynthesis of Nonribosomal Peptides"

    Article Title: ClbP Is a Prototype of a Peptidase Subgroup Involved in Biosynthesis of Nonribosomal Peptides

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.221960

    Key residues in the ClbP active site. A , residues of ClbP located in close vicinity of the binding site. Hydrogen bonds are indicated by pink dashed lines , and the water molecules are indicated by W . Carbon atoms are colored green , oxygen atoms are colored red, and nitrogen atoms are colored blue. B , overlay of penicillin-recognizing motifs of ClbP, Pab87 peptidase, and AmpC β-lactamase. Carbon atoms of ClbP, Pab87 peptidase, and AmpC β-lactamase are in green , yellow , and gray , respectively. The motifs S xx K, Y x (N/S/T), and (K/H)(S/T/G)G are numbered in red , blue , and green , respectively, and residue numbering is according to the ClbP sequence. C and D , phenotypic analysis of pks island activity in E. coli DH10B/pBAC pks deleted or not for the clbP gene and trans-complemented with ClbP WT protein or the S95A, K98T, or Y186G active site ClbP mutants. HeLa cells were infected 4 h with 100 E. coli per cell, and then the cells were washed and incubated with gentamicin for 4–72 h. C , anti-5His Western blot analysis of the expression of His-tagged WT and active site ClbP mutants in E. coli DH10B/pBAC pks Δ clbP after infection. D , host histone H2AX Ser-139 phosphorylation (γH2AX) indicative of DNA double-strand breaks was assayed by confocal immunofluorescence 4 h after infection. DNA and γH2AX are pseudocolored in blue and red , respectively ( bars , 20 μm). Cell swelling (megalocytosis) was observed following Giemsa staining 72 h after infection ( bars , 50 μm). G 2 cell cycle arrest following DNA damage was assessed by flow cytometric DNA content analysis, 48 h after infection.
    Figure Legend Snippet: Key residues in the ClbP active site. A , residues of ClbP located in close vicinity of the binding site. Hydrogen bonds are indicated by pink dashed lines , and the water molecules are indicated by W . Carbon atoms are colored green , oxygen atoms are colored red, and nitrogen atoms are colored blue. B , overlay of penicillin-recognizing motifs of ClbP, Pab87 peptidase, and AmpC β-lactamase. Carbon atoms of ClbP, Pab87 peptidase, and AmpC β-lactamase are in green , yellow , and gray , respectively. The motifs S xx K, Y x (N/S/T), and (K/H)(S/T/G)G are numbered in red , blue , and green , respectively, and residue numbering is according to the ClbP sequence. C and D , phenotypic analysis of pks island activity in E. coli DH10B/pBAC pks deleted or not for the clbP gene and trans-complemented with ClbP WT protein or the S95A, K98T, or Y186G active site ClbP mutants. HeLa cells were infected 4 h with 100 E. coli per cell, and then the cells were washed and incubated with gentamicin for 4–72 h. C , anti-5His Western blot analysis of the expression of His-tagged WT and active site ClbP mutants in E. coli DH10B/pBAC pks Δ clbP after infection. D , host histone H2AX Ser-139 phosphorylation (γH2AX) indicative of DNA double-strand breaks was assayed by confocal immunofluorescence 4 h after infection. DNA and γH2AX are pseudocolored in blue and red , respectively ( bars , 20 μm). Cell swelling (megalocytosis) was observed following Giemsa staining 72 h after infection ( bars , 50 μm). G 2 cell cycle arrest following DNA damage was assessed by flow cytometric DNA content analysis, 48 h after infection.

    Techniques Used: Binding Assay, Sequencing, Activity Assay, Infection, Incubation, Western Blot, Expressing, Immunofluorescence, Staining, Flow Cytometry

    5) Product Images from "Synergism and the mechanism of action of the combination of α-mangostin isolated from Garcinia mangostana L. and oxacillin against an oxacillin-resistant Staphylococcus saprophyticus"

    Article Title: Synergism and the mechanism of action of the combination of α-mangostin isolated from Garcinia mangostana L. and oxacillin against an oxacillin-resistant Staphylococcus saprophyticus

    Journal: BMC Microbiology

    doi: 10.1186/s12866-016-0814-4

    The inhibitory activity of α-mangostin against β-lactamase in hydrolysing benzylpenicillin. β-lactamase used from E. cloacae ; CON = control (no testing agent); AMT(1) = 1 μg/ml α-mangostin. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Scheffe’s test, p
    Figure Legend Snippet: The inhibitory activity of α-mangostin against β-lactamase in hydrolysing benzylpenicillin. β-lactamase used from E. cloacae ; CON = control (no testing agent); AMT(1) = 1 μg/ml α-mangostin. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Scheffe’s test, p

    Techniques Used: Activity Assay

    6) Product Images from "Copper Influences the Antibacterial Outcomes of a β-Lactamase-Activated Prochelator against Drug-Resistant Bacteria"

    Article Title: Copper Influences the Antibacterial Outcomes of a β-Lactamase-Activated Prochelator against Drug-Resistant Bacteria

    Journal: ACS infectious diseases

    doi: 10.1021/acsinfecdis.8b00037

    Prochelator PcephPT inhibits growth of β-lactamase-expressing E. coli .
    Figure Legend Snippet: Prochelator PcephPT inhibits growth of β-lactamase-expressing E. coli .

    Techniques Used: Expressing

    Ability of PcephPT to kill MG1655 bacteria depends on both the presence of β-lactamase and availability of Cu.
    Figure Legend Snippet: Ability of PcephPT to kill MG1655 bacteria depends on both the presence of β-lactamase and availability of Cu.

    Techniques Used:

    Prochelator PcephPT is selectively turned over by bacteria that produce β-lactamase.
    Figure Legend Snippet: Prochelator PcephPT is selectively turned over by bacteria that produce β-lactamase.

    Techniques Used:

    7) Product Images from "Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria"

    Article Title: Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.72.1.880-889.2006

    Immobilization of enzymes on the surface of lactococcal GEM particles. (a) Relative α-amylase activities on GEM particles incubated with culture medium containing soluble AmyL, PA (PA3), or α-PA. (b) Relative α-amylase and β-lactamase
    Figure Legend Snippet: Immobilization of enzymes on the surface of lactococcal GEM particles. (a) Relative α-amylase activities on GEM particles incubated with culture medium containing soluble AmyL, PA (PA3), or α-PA. (b) Relative α-amylase and β-lactamase

    Techniques Used: Incubation

    8) Product Images from "Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein"

    Article Title: Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.45.12.3279-3286.2001

    Purified β-lactamase proteins. The six β-lactamase proteins were purified to > 90% homogeneity, as determined by SDS-PAGE. The values on the molecular weight ladder (MWL) are expressed in units of kilodaltons.
    Figure Legend Snippet: Purified β-lactamase proteins. The six β-lactamase proteins were purified to > 90% homogeneity, as determined by SDS-PAGE. The values on the molecular weight ladder (MWL) are expressed in units of kilodaltons.

    Techniques Used: Purification, SDS Page, Molecular Weight

    ). Closed squares, protein kinase C peptide; open squares, cyclic BP46-51 peptide; closed circles, reduced BP46-51 peptide; open circles, BP41-50 peptide. Each datum point is the average for two independent experiments. (B) Inhibition assay of TEM-1 β-lactamase by wild-type BLIP. A K i of 0.23 nM was determined by using a nonlinear regression fit as described above.
    Figure Legend Snippet: ). Closed squares, protein kinase C peptide; open squares, cyclic BP46-51 peptide; closed circles, reduced BP46-51 peptide; open circles, BP41-50 peptide. Each datum point is the average for two independent experiments. (B) Inhibition assay of TEM-1 β-lactamase by wild-type BLIP. A K i of 0.23 nM was determined by using a nonlinear regression fit as described above.

    Techniques Used: Inhibition, Transmission Electron Microscopy

    9) Product Images from "Genetic Selection for Enhanced Folding In Vivo Targets the Cys14-Cys38 Disulfide Bond in Bovine Pancreatic Trypsin Inhibitor"

    Article Title: Genetic Selection for Enhanced Folding In Vivo Targets the Cys14-Cys38 Disulfide Bond in Bovine Pancreatic Trypsin Inhibitor

    Journal: Antioxidants & Redox Signaling

    doi: 10.1089/ars.2010.3712

    The stabilities of single disulfide-bonded BPTI species in vitro correlate with the level of antibiotic resistance in the β-lactamase system in vivo
    Figure Legend Snippet: The stabilities of single disulfide-bonded BPTI species in vitro correlate with the level of antibiotic resistance in the β-lactamase system in vivo

    Techniques Used: In Vitro, In Vivo

    Insertion of BPTI into β-lactamase leads to a massive drop in antibiotic resistance compared to cells expressing the unmodified enzyme. Mid-log phase cells of E. coli MG1655 Δ ampC expressing TEM1-β-lactamase (bla WT) or TEM1-β-lactamase
    Figure Legend Snippet: Insertion of BPTI into β-lactamase leads to a massive drop in antibiotic resistance compared to cells expressing the unmodified enzyme. Mid-log phase cells of E. coli MG1655 Δ ampC expressing TEM1-β-lactamase (bla WT) or TEM1-β-lactamase

    Techniques Used: Expressing

    Directed evolution of proteins using the modified protein stability increased by directed evolution and the β-lactamase tripartite fusion systems. The test protein BPTI (shown in orange) is inserted into either the minor coat protein of the fd
    Figure Legend Snippet: Directed evolution of proteins using the modified protein stability increased by directed evolution and the β-lactamase tripartite fusion systems. The test protein BPTI (shown in orange) is inserted into either the minor coat protein of the fd

    Techniques Used: Modification

    Selecting for BPTI variants that lead to increased antibiotic resistance in the β-lactamase system
    Figure Legend Snippet: Selecting for BPTI variants that lead to increased antibiotic resistance in the β-lactamase system

    Techniques Used:

    Increased destabilization of the Cys14-Cys38 disulfide bond in vitro correlates with increased antibiotic resistance in the β - lactamase system in vivo. Different variants of BPTI were inserted into β-lactamase via flexible linkers. The
    Figure Legend Snippet: Increased destabilization of the Cys14-Cys38 disulfide bond in vitro correlates with increased antibiotic resistance in the β - lactamase system in vivo. Different variants of BPTI were inserted into β-lactamase via flexible linkers. The

    Techniques Used: In Vitro, In Vivo

    In BPTI variants that can only form a single disulfide bond, the stability of the disulfide bond in vitro correlates with the level of antibiotic resistance in the β - lactamase system in vivo. (A) Three variants of BPTI representing single disulfide-bonded
    Figure Legend Snippet: In BPTI variants that can only form a single disulfide bond, the stability of the disulfide bond in vitro correlates with the level of antibiotic resistance in the β - lactamase system in vivo. (A) Three variants of BPTI representing single disulfide-bonded

    Techniques Used: In Vitro, In Vivo

    Substitution of Cys17 in hG-CSF with serine leads to an increase in antibiotic resistance as part of the β-lactamase system. WT hG-CSF, which contains two disulfide bonds (Cys36-Cys42 and Cys64-Cys74) and one unpaired cysteine (Cys17), is prone
    Figure Legend Snippet: Substitution of Cys17 in hG-CSF with serine leads to an increase in antibiotic resistance as part of the β-lactamase system. WT hG-CSF, which contains two disulfide bonds (Cys36-Cys42 and Cys64-Cys74) and one unpaired cysteine (Cys17), is prone

    Techniques Used:

    10) Product Images from "Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿ †"

    Article Title: Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿ †

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.05363-11

    HPLC analysis of the turnover of the cephalosporins cephaloridine (A), cephalothin (B), and cefazolin (C) by EstU1 (green) and class C β-lactamase of E. cloacae (red). The chromatogram of cephalosporins is shown as a black line. The retention
    Figure Legend Snippet: HPLC analysis of the turnover of the cephalosporins cephaloridine (A), cephalothin (B), and cefazolin (C) by EstU1 (green) and class C β-lactamase of E. cloacae (red). The chromatogram of cephalosporins is shown as a black line. The retention

    Techniques Used: High Performance Liquid Chromatography

    11) Product Images from "Hydrolysis Spectrum Extension of CMY-2-Like ?-Lactamases Resulting from Structural Alteration in the Y-X-N Loop"

    Article Title: Hydrolysis Spectrum Extension of CMY-2-Like ?-Lactamases Resulting from Structural Alteration in the Y-X-N Loop

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.05630-11

    Representation of the secondary structures surrounding the amino acid at position 148. Left panel, Native AmpC β-lactamase of C. freundii GN346 (PDB 1RGY ); Right panel, AmpC variant of the AmpC β-lactamase of C. freundii GN346 presenting
    Figure Legend Snippet: Representation of the secondary structures surrounding the amino acid at position 148. Left panel, Native AmpC β-lactamase of C. freundii GN346 (PDB 1RGY ); Right panel, AmpC variant of the AmpC β-lactamase of C. freundii GN346 presenting

    Techniques Used: Variant Assay

    12) Product Images from "Design and Exploration of Novel Boronic Acid Inhibitors Reveals Important Interactions with a Clavulanic Acid-Resistant Sulfhydryl-Variable (SHV) \u03b2-Lactamase"

    Article Title: Design and Exploration of Novel Boronic Acid Inhibitors Reveals Important Interactions with a Clavulanic Acid-Resistant Sulfhydryl-Variable (SHV) \u03b2-Lactamase

    Journal: Journal of medicinal chemistry

    doi: 10.1021/jm301490d

    The top figure shows the wild-type enzyme with Ser-130 at a χ angle of −144° where proton shuffling can occur to allow breakdown of ampicillin by the β-lactamase. The bottom figure shows the movement of the hydroxyl group
    Figure Legend Snippet: The top figure shows the wild-type enzyme with Ser-130 at a χ angle of −144° where proton shuffling can occur to allow breakdown of ampicillin by the β-lactamase. The bottom figure shows the movement of the hydroxyl group

    Techniques Used:

    Kinetics of K234R with β-Lactamase Inhibitors
    Figure Legend Snippet: Kinetics of K234R with β-Lactamase Inhibitors

    Techniques Used:

    13) Product Images from "Synergistic activity and mechanism of action of Stephania suberosa Forman extract and ampicillin combination against ampicillin-resistant Staphylococcus aureus"

    Article Title: Synergistic activity and mechanism of action of Stephania suberosa Forman extract and ampicillin combination against ampicillin-resistant Staphylococcus aureus

    Journal: Journal of Biomedical Science

    doi: 10.1186/s12929-014-0090-2

    The inhibitory activity of SSE against β-lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; Con = control (no testing agent); SSE(1) = SSE at 1 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey’s HSD, p
    Figure Legend Snippet: The inhibitory activity of SSE against β-lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; Con = control (no testing agent); SSE(1) = SSE at 1 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey’s HSD, p

    Techniques Used: Activity Assay

    14) Product Images from "A Mutation in the Ebola Virus Envelope Glycoprotein Restricts Viral Entry in a Host Species- and Cell-Type-Specific Manner"

    Article Title: A Mutation in the Ebola Virus Envelope Glycoprotein Restricts Viral Entry in a Host Species- and Cell-Type-Specific Manner

    Journal: Journal of Virology

    doi: 10.1128/JVI.01598-12

    Impacts of mutations to hydrophobic residues near F88 on entry into different cell types. (A) Equivalent amounts, based on β-lactamase activity, of GP-wt, GP-F88A, GP-L111A, GP-I113A, GP-L122A, GP-F159A, and GP-F225A VLPs were generated and tested
    Figure Legend Snippet: Impacts of mutations to hydrophobic residues near F88 on entry into different cell types. (A) Equivalent amounts, based on β-lactamase activity, of GP-wt, GP-F88A, GP-L111A, GP-I113A, GP-L122A, GP-F159A, and GP-F225A VLPs were generated and tested

    Techniques Used: Activity Assay, Generated

    GP-F88A is permissive for entry into mouse peritoneal cells, but not human dendritic cells. Equivalent amounts of VLPs, as determined by assaying the total β-lactamase activity of purified VLP preparations possessing wt GP [GP (wt)], GP-F88A [GP
    Figure Legend Snippet: GP-F88A is permissive for entry into mouse peritoneal cells, but not human dendritic cells. Equivalent amounts of VLPs, as determined by assaying the total β-lactamase activity of purified VLP preparations possessing wt GP [GP (wt)], GP-F88A [GP

    Techniques Used: Activity Assay, Purification

    Mouse IC-21 macrophages are susceptible to entry by GP-F88A VLPs and permit infection with GP-F88A-pseudotyped VSV. (A and B) Equivalent amounts, based on β-lactamase activity, of GP-wt and GP-F88A VLPs were tested for entry into RAW cells (A)
    Figure Legend Snippet: Mouse IC-21 macrophages are susceptible to entry by GP-F88A VLPs and permit infection with GP-F88A-pseudotyped VSV. (A and B) Equivalent amounts, based on β-lactamase activity, of GP-wt and GP-F88A VLPs were tested for entry into RAW cells (A)

    Techniques Used: Infection, Activity Assay

    15) Product Images from "Moraxella catarrhalis Outer Membrane Vesicles Carry ?-Lactamase and Promote Survival of Streptococcus pneumoniae and Haemophilus influenzae by Inactivating Amoxicillin ▿"

    Article Title: Moraxella catarrhalis Outer Membrane Vesicles Carry ?-Lactamase and Promote Survival of Streptococcus pneumoniae and Haemophilus influenzae by Inactivating Amoxicillin ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01772-10

    M. catarrhalis β-lactamase-containing OMVs protect S. pneumoniae (pneumococci [Pnc]) from being killed by amoxicillin. Amoxicillin-susceptible S. pneumoniae ATCC 6303 (10 6 CFU/ml) was grown with amoxicillin (1 μg/ml) preincubated with either 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). OMVs were isolated from M. catarrhalis KR526 (β-lac + ) and Bc5 (β-lac − ). Growth was expressed either as relative growth compared to starting concentrations measured as absorbance (OD 600 ) (A and C) or as numbers of CFU (B). The data are presented as means and the standard errors of at least three independent experiments. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.
    Figure Legend Snippet: M. catarrhalis β-lactamase-containing OMVs protect S. pneumoniae (pneumococci [Pnc]) from being killed by amoxicillin. Amoxicillin-susceptible S. pneumoniae ATCC 6303 (10 6 CFU/ml) was grown with amoxicillin (1 μg/ml) preincubated with either 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). OMVs were isolated from M. catarrhalis KR526 (β-lac + ) and Bc5 (β-lac − ). Growth was expressed either as relative growth compared to starting concentrations measured as absorbance (OD 600 ) (A and C) or as numbers of CFU (B). The data are presented as means and the standard errors of at least three independent experiments. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.

    Techniques Used: Isolation

    Vesicles from β-lactamase-positive M. catarrhalis protect NTHi against amoxicillin. Amoxicillin-susceptible NTHi 772 (A to C) or Hib KR124 (D) (10 7 CFU/ml) was grown with amoxicillin (2 μg/ml) that had been preincubated with either 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). OMVs were isolated from M. catarrhalis KR526 and Bc5, which were β-lactamase positive and negative, respectively. Growth was expressed either as relative growth compared to starting concentrations measured as absorbance at OD 600 (A and C) or as numbers of CFU (B and D). The results are shown as means and SEMs of at least three independent experiments. *, P ≤ 0.05; ***, P ≤ 0.001.
    Figure Legend Snippet: Vesicles from β-lactamase-positive M. catarrhalis protect NTHi against amoxicillin. Amoxicillin-susceptible NTHi 772 (A to C) or Hib KR124 (D) (10 7 CFU/ml) was grown with amoxicillin (2 μg/ml) that had been preincubated with either 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). OMVs were isolated from M. catarrhalis KR526 and Bc5, which were β-lactamase positive and negative, respectively. Growth was expressed either as relative growth compared to starting concentrations measured as absorbance at OD 600 (A and C) or as numbers of CFU (B and D). The results are shown as means and SEMs of at least three independent experiments. *, P ≤ 0.05; ***, P ≤ 0.001.

    Techniques Used: Isolation

    M. catarrhalis OMVs contain enzymatically active β-lactamase. (A) The β-lactamase contents of whole-cell lysates and OMVs from four different M. catarrhalis strains were analyzed. OMVs contained approximately the same β-lactamase content as whole-cell lysates. (B) The β-lactamase enzyme was found on the inside of the OMVs. The β-lactamase activity was quantified by the ability of the enzyme to hydrolyze the β-lactam nitrocefin, leading to a change in absorbance from OD 380 to OD 485 , as determined by spectrophotometry. β-Lactamase enzyme or OMVs were treated with proteinase K (100 μg/ml) and/or saponin (0.2%), and enzyme content was subsequently determined. As a negative control, OMVs were first treated with saponin, followed by proteinase K. In panel A, the β-lactamase content was expressed as the number of moles nitrocefin hydrolyzed per minute per mg OMVs. In panel B, β-lactamase activity was defined as the percentage of enzyme activity of OMV in the absence of amoxicillin that was set to 100%. Data shown are means and SEM of at least three independent experiments.
    Figure Legend Snippet: M. catarrhalis OMVs contain enzymatically active β-lactamase. (A) The β-lactamase contents of whole-cell lysates and OMVs from four different M. catarrhalis strains were analyzed. OMVs contained approximately the same β-lactamase content as whole-cell lysates. (B) The β-lactamase enzyme was found on the inside of the OMVs. The β-lactamase activity was quantified by the ability of the enzyme to hydrolyze the β-lactam nitrocefin, leading to a change in absorbance from OD 380 to OD 485 , as determined by spectrophotometry. β-Lactamase enzyme or OMVs were treated with proteinase K (100 μg/ml) and/or saponin (0.2%), and enzyme content was subsequently determined. As a negative control, OMVs were first treated with saponin, followed by proteinase K. In panel A, the β-lactamase content was expressed as the number of moles nitrocefin hydrolyzed per minute per mg OMVs. In panel B, β-lactamase activity was defined as the percentage of enzyme activity of OMV in the absence of amoxicillin that was set to 100%. Data shown are means and SEM of at least three independent experiments.

    Techniques Used: Activity Assay, Spectrophotometry, Negative Control

    Amoxicillin-resistant M. catarrhalis strains produce OMV containing β-lactamase. (A) Eight M. catarrhalis strains out of 10 were positive for the bro gene (522 bp), as revealed by PCR analysis. bro alleles were not found in strains KR395 and Bc5. (B) M. catarrhalis OMVs and total bacterial lysates (10 μg each) were subjected to SDS-PAGE (left), followed by detection of β-lactamase (35 kDa) by Western blotting (right). Lysates of whole M. catarrhalis RH4 and KR395 bacteria were used as positive and negative controls, respectively. Recombinant RH4 β-lactamase (0.6 μg) was also included. The upper band represents the recombinant protein at a size of 37.7 kDa. The lower band most likely results from N-terminal degradation. The His tag located at the C-terminal end was not affected by degradation, since it was possible to purify the recombinant β-lactamase using affinity chromatography. (C) Flow cytometry using antiserum raised against recombinant RH4 β-lactamase. The arrow shows a positive shift with β-lactamase containing OMV KR526, whereas OMVs from Bc5 were negative. OMVs without the β-lactamase antiserum (black) were compared to OMVs incubated with β-lactamase antiserum (white). (D) Gold-labeled anti-β-lactamase antibodies confirmed the presence of β-lactamase by TEM.
    Figure Legend Snippet: Amoxicillin-resistant M. catarrhalis strains produce OMV containing β-lactamase. (A) Eight M. catarrhalis strains out of 10 were positive for the bro gene (522 bp), as revealed by PCR analysis. bro alleles were not found in strains KR395 and Bc5. (B) M. catarrhalis OMVs and total bacterial lysates (10 μg each) were subjected to SDS-PAGE (left), followed by detection of β-lactamase (35 kDa) by Western blotting (right). Lysates of whole M. catarrhalis RH4 and KR395 bacteria were used as positive and negative controls, respectively. Recombinant RH4 β-lactamase (0.6 μg) was also included. The upper band represents the recombinant protein at a size of 37.7 kDa. The lower band most likely results from N-terminal degradation. The His tag located at the C-terminal end was not affected by degradation, since it was possible to purify the recombinant β-lactamase using affinity chromatography. (C) Flow cytometry using antiserum raised against recombinant RH4 β-lactamase. The arrow shows a positive shift with β-lactamase containing OMV KR526, whereas OMVs from Bc5 were negative. OMVs without the β-lactamase antiserum (black) were compared to OMVs incubated with β-lactamase antiserum (white). (D) Gold-labeled anti-β-lactamase antibodies confirmed the presence of β-lactamase by TEM.

    Techniques Used: Polymerase Chain Reaction, SDS Page, Western Blot, Recombinant, Affinity Chromatography, Flow Cytometry, Cytometry, Incubation, Labeling, Transmission Electron Microscopy

    β-Lactamase-carrying M. catarrhalis OMVs hydrolyze amoxicillin. (A) Amoxicillin (AMX)-induced killing at 1.25 to 10 μg/ml amoxicillin was gradually reduced with increasing concentrations (0 to 50 μg/ml) of β-lactamase-containing OMVs. Amoxicillin concentrations were determined by measuring inhibitory growth zones of the β-lactam-susceptible bacterium Sarcina lutea . (B) β-Lactamase-positive and -negative M. catarrhalis OMVs at 25 μg/ml were incubated with increasing amoxicillin concentrations. The data are presented as means and SEMs of at least three independent experiments. β-lac + and β-lac − , β-lactamase positive and negative, respectively. ***, P ≤ 0.001.
    Figure Legend Snippet: β-Lactamase-carrying M. catarrhalis OMVs hydrolyze amoxicillin. (A) Amoxicillin (AMX)-induced killing at 1.25 to 10 μg/ml amoxicillin was gradually reduced with increasing concentrations (0 to 50 μg/ml) of β-lactamase-containing OMVs. Amoxicillin concentrations were determined by measuring inhibitory growth zones of the β-lactam-susceptible bacterium Sarcina lutea . (B) β-Lactamase-positive and -negative M. catarrhalis OMVs at 25 μg/ml were incubated with increasing amoxicillin concentrations. The data are presented as means and SEMs of at least three independent experiments. β-lac + and β-lac − , β-lactamase positive and negative, respectively. ***, P ≤ 0.001.

    Techniques Used: Incubation

    β-Lactamase-positive M. catarrhalis OMVs protect amoxicillin-susceptible M. catarrhalis strains from being killed by amoxicillin. β-Lactamase-susceptible M. catarrhalis KR935 (10 7 CFU/ml) was grown with amoxicillin (1 μg/ml) that had been preincubated in the presence of 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). β-Lactamase-positive and -negative OMVs were isolated from M. catarrhalis KR526 (β-lac + ) and Bc5 (β-lac − ), respectively. Growth was expressed either as relative growth compared to starting concentrations measured as absorbance (OD 600 ) (A and C) or as numbers of CFU (B). Mean values and SEMs of at least three independent experiments are shown. *, P ≤ 0.05.
    Figure Legend Snippet: β-Lactamase-positive M. catarrhalis OMVs protect amoxicillin-susceptible M. catarrhalis strains from being killed by amoxicillin. β-Lactamase-susceptible M. catarrhalis KR935 (10 7 CFU/ml) was grown with amoxicillin (1 μg/ml) that had been preincubated in the presence of 25 μg/ml β-lactamase-positive OMVs (A and B) or β-lactamase-negative OMVs (B and C). β-Lactamase-positive and -negative OMVs were isolated from M. catarrhalis KR526 (β-lac + ) and Bc5 (β-lac − ), respectively. Growth was expressed either as relative growth compared to starting concentrations measured as absorbance (OD 600 ) (A and C) or as numbers of CFU (B). Mean values and SEMs of at least three independent experiments are shown. *, P ≤ 0.05.

    Techniques Used: Isolation

    16) Product Images from "Horizontal Transfer of blaCMY-Bearing Plasmids among Clinical Escherichia coli and Klebsiella pneumoniae Isolates and Emergence of Cefepime-Hydrolyzing CMY-19"

    Article Title: Horizontal Transfer of blaCMY-Bearing Plasmids among Clinical Escherichia coli and Klebsiella pneumoniae Isolates and Emergence of Cefepime-Hydrolyzing CMY-19

    Journal:

    doi: 10.1128/AAC.50.2.534-541.2006

    Plasmid profiles and Southern hybridization. (A) Plasmid profiles of clinical isolates and their tranconjugants; (B) hybridization with the probe specific for the CMY-1- and MOX-1-type β-lactamase gene. Lanes: M, HindIII-digested DNA marker; 1,
    Figure Legend Snippet: Plasmid profiles and Southern hybridization. (A) Plasmid profiles of clinical isolates and their tranconjugants; (B) hybridization with the probe specific for the CMY-1- and MOX-1-type β-lactamase gene. Lanes: M, HindIII-digested DNA marker; 1,

    Techniques Used: Plasmid Preparation, Hybridization, Marker

    Plasmid patterns after restriction enzyme digestion and Southern hybridization. (A) SacI-digested plasmid DNAs prepared from the representative transconjugants; (B) hybridization patterns with the probe specific for CMY-1- and MOX-1-type β-lactamase
    Figure Legend Snippet: Plasmid patterns after restriction enzyme digestion and Southern hybridization. (A) SacI-digested plasmid DNAs prepared from the representative transconjugants; (B) hybridization patterns with the probe specific for CMY-1- and MOX-1-type β-lactamase

    Techniques Used: Plasmid Preparation, Hybridization

    17) Product Images from "Massively Parallel Delivery of Large-Sized Cargo into Mammalian Cells with Light Pulses"

    Article Title: Massively Parallel Delivery of Large-Sized Cargo into Mammalian Cells with Light Pulses

    Journal: Nature methods

    doi: 10.1038/nmeth.3357

    BLAST delivered β-lactamase enzyme is functional inside NHDFs
    Figure Legend Snippet: BLAST delivered β-lactamase enzyme is functional inside NHDFs

    Techniques Used: Functional Assay

    18) Product Images from "Synergy and Mode of Action of Ceftazidime plus Quercetin or Luteolin on Streptococcus pyogenes"

    Article Title: Synergy and Mode of Action of Ceftazidime plus Quercetin or Luteolin on Streptococcus pyogenes

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2015/759459

    The inhibitory activity of luteolin, quercetin, and ceftazidime against β -lactamase in hydrolyzing benzylpenicillin. β -lactamase used from E. cloacae ; Con = control (no testing agent), Cef (0.25) = 0.25 μ g mL −1 , Que (64) = quercetin 64 μ g mL −1 , and Lut (64) = luteolin 64 μ g mL −1 . The graph shows the remaining benzylpenicillin at the same time. The research was executed in three studies, and all graphs are displayed as mean ± SEM. Means sharing the same superscript are not significantly different from each other (Scheffe's, P
    Figure Legend Snippet: The inhibitory activity of luteolin, quercetin, and ceftazidime against β -lactamase in hydrolyzing benzylpenicillin. β -lactamase used from E. cloacae ; Con = control (no testing agent), Cef (0.25) = 0.25 μ g mL −1 , Que (64) = quercetin 64 μ g mL −1 , and Lut (64) = luteolin 64 μ g mL −1 . The graph shows the remaining benzylpenicillin at the same time. The research was executed in three studies, and all graphs are displayed as mean ± SEM. Means sharing the same superscript are not significantly different from each other (Scheffe's, P

    Techniques Used: Activity Assay

    19) Product Images from "Design and Exploration of Novel Boronic Acid Inhibitors Reveals Important Interactions with a Clavulanic Acid-Resistant Sulfhydryl-Variable (SHV) \u03b2-Lactamase"

    Article Title: Design and Exploration of Novel Boronic Acid Inhibitors Reveals Important Interactions with a Clavulanic Acid-Resistant Sulfhydryl-Variable (SHV) \u03b2-Lactamase

    Journal: Journal of medicinal chemistry

    doi: 10.1021/jm301490d

    The top figure shows the wild-type enzyme with Ser-130 at a χ angle of −144° where proton shuffling can occur to allow breakdown of ampicillin by the β-lactamase. The bottom figure shows the movement of the hydroxyl group
    Figure Legend Snippet: The top figure shows the wild-type enzyme with Ser-130 at a χ angle of −144° where proton shuffling can occur to allow breakdown of ampicillin by the β-lactamase. The bottom figure shows the movement of the hydroxyl group

    Techniques Used:

    Kinetics of K234R with β-Lactamase Inhibitors
    Figure Legend Snippet: Kinetics of K234R with β-Lactamase Inhibitors

    Techniques Used:

    20) Product Images from "The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone"

    Article Title: The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.186.11.3621-3630.2004

    HopPtoV is secreted in culture via the DC3000 TTSS in an ShcV-dependent manner. (A) Cultures of wild-type (WT) P. syringae pv. tomato DC3000 and a DC3000 TTSS-defective mutant ( hrcC ) were grown in hrp -inducing medium and separated into cell-bound (C) and supernatant (S) fractions. The cell-bound and supernatant fractions were concentrated 13.3-fold and 133-fold, respectively, relative to the initial culture volumes. The samples were separated by SDS-PAGE and immunoblotted. HopPtoV and β-lactamase, which was used as a lysis control, were detected with anti-HA and anti-β-lactamase antibodies, respectively. Wild-type DC3000 strains containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) secreted HopPtoV-HA into the supernatant fraction. However, much more HopPtoV-HA was found in the supernatant fractions from DC3000(pLN517), which overexpressed ShcV. (B) Cultures of DC3000 shcV (UNL120) and hopPtoV (UNL125) insertion mutants containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) were grown in hrp -inducing medium, separated into cell-bound (C) and supernatant (S) fractions, and separated by SDS-PAGE. Immunoblotted proteins were detected with anti-HA or anti-NPTII antibodies. Plasmid-encoded NPTII should remain cell bound and was used as a lysis control. The shcV mutant UNL120 was unable to secrete HopPtoV-HA unless shcV was also provided in trans .
    Figure Legend Snippet: HopPtoV is secreted in culture via the DC3000 TTSS in an ShcV-dependent manner. (A) Cultures of wild-type (WT) P. syringae pv. tomato DC3000 and a DC3000 TTSS-defective mutant ( hrcC ) were grown in hrp -inducing medium and separated into cell-bound (C) and supernatant (S) fractions. The cell-bound and supernatant fractions were concentrated 13.3-fold and 133-fold, respectively, relative to the initial culture volumes. The samples were separated by SDS-PAGE and immunoblotted. HopPtoV and β-lactamase, which was used as a lysis control, were detected with anti-HA and anti-β-lactamase antibodies, respectively. Wild-type DC3000 strains containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) secreted HopPtoV-HA into the supernatant fraction. However, much more HopPtoV-HA was found in the supernatant fractions from DC3000(pLN517), which overexpressed ShcV. (B) Cultures of DC3000 shcV (UNL120) and hopPtoV (UNL125) insertion mutants containing plasmid pLN127 (p hopPtoV-ha ) or pLN517 (p shcV/hopPtoV-ha ) were grown in hrp -inducing medium, separated into cell-bound (C) and supernatant (S) fractions, and separated by SDS-PAGE. Immunoblotted proteins were detected with anti-HA or anti-NPTII antibodies. Plasmid-encoded NPTII should remain cell bound and was used as a lysis control. The shcV mutant UNL120 was unable to secrete HopPtoV-HA unless shcV was also provided in trans .

    Techniques Used: Mutagenesis, SDS Page, Lysis, Plasmid Preparation

    21) Product Images from "Non-canonical autophagy in dendritic cells restricts cross-presentation and anti-tumor immunity"

    Article Title: Non-canonical autophagy in dendritic cells restricts cross-presentation and anti-tumor immunity

    Journal: bioRxiv

    doi: 10.1101/789867

    Rubcn -/- DCs exhibit increased phagosome-to-cytosol escape and proteasome-mediated generation of peptides. Bone marrow-derived dendritic cells (DCs) were generated from Rubcn +/+ (black) and Rubcn -/- (red) mice in vitro with FLT3-L for 7 days. ( A-B ) DCs were loaded with 1 µm CCF4, then co-cultured with apoptotic B16-OVA in the absence or presence of β-lactamase (2 mg/ml) for 90 or 180 minutes at 4°C or 37°C. Uncleaved CCF4 was measured by flow cytometry at an emission of 535 nm, and cleaved CCF4 was measured at an emission of 450 nm. Ratios of 450 nm : 535 nm was calculated by dividing the 450 nm MFI by the 535 nm MFI from a single sample. ( C ) DCs were pre-treated with vehicle or chloroquine (CQ) at 1 or 10 µM for 2 hours and then co-cultured with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( D ) DCs were pre-treated with vehicle or MG-132 at 1 or 10 µM for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( E ) DCs were pre-treated with vehicle or Brefeldin A at 3 µg/ml for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. Data are expressed as mean ± SEM. No less than two independent experiments were performed, with 3-5 replicates per condition. Significance was calculated using 2-way ANOVA (*p
    Figure Legend Snippet: Rubcn -/- DCs exhibit increased phagosome-to-cytosol escape and proteasome-mediated generation of peptides. Bone marrow-derived dendritic cells (DCs) were generated from Rubcn +/+ (black) and Rubcn -/- (red) mice in vitro with FLT3-L for 7 days. ( A-B ) DCs were loaded with 1 µm CCF4, then co-cultured with apoptotic B16-OVA in the absence or presence of β-lactamase (2 mg/ml) for 90 or 180 minutes at 4°C or 37°C. Uncleaved CCF4 was measured by flow cytometry at an emission of 535 nm, and cleaved CCF4 was measured at an emission of 450 nm. Ratios of 450 nm : 535 nm was calculated by dividing the 450 nm MFI by the 535 nm MFI from a single sample. ( C ) DCs were pre-treated with vehicle or chloroquine (CQ) at 1 or 10 µM for 2 hours and then co-cultured with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( D ) DCs were pre-treated with vehicle or MG-132 at 1 or 10 µM for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( E ) DCs were pre-treated with vehicle or Brefeldin A at 3 µg/ml for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. Data are expressed as mean ± SEM. No less than two independent experiments were performed, with 3-5 replicates per condition. Significance was calculated using 2-way ANOVA (*p

    Techniques Used: Derivative Assay, Generated, Mouse Assay, In Vitro, Cell Culture, Flow Cytometry, Expressing

    22) Product Images from "Growth phenotypes of Pseudomonas aeruginosa lasR mutants adapted to the airways of cystic fibrosis patients"

    Article Title: Growth phenotypes of Pseudomonas aeruginosa lasR mutants adapted to the airways of cystic fibrosis patients

    Journal: Molecular microbiology

    doi: 10.1111/j.1365-2958.2007.05678.x

    Inactivation of lasR confers increased β-lactamase activity and β-lactam tolerance. A. Ceftazidime-resistant colonies emerged in a lawn of CF416 or CF416L1 cells on LB agar with 20 µg ml −1 of ceftazidime (Petri dish upper and lower halves), but only the lasR mutant lawns yielded small partially resistant colonies whose growth is inhibited by the addition of 8 µg ml −1 of the β-lactamase inhibitor tazobactam (Petri dish lower halves). Photographs were taken after incubation overnight at 37°C followed by 3 days at 22°C. B. β-Lactamase activity in culture supernatants, measured as a change in optical density at 490 nm (ΔA 490 ) after addition of the chromogenic substrate nitrocefin. Cases with no change are marked with an asterisk. Values are the average of three technical replicates, and error bars show standard deviations. Equivalent results were obtained using a qualitative whole-culture assay. C. β-Lactamase activity in whole cultures of CF416 and CF416 lasR ::Gm. Values are the averages of three cultures, and error bars show standard error of the mean. One unit of β-lactamase is defined as 1 µmol of nitrocefin hydrolysed per min per mg of total protein. D. Reduced killing by ceftazidime of the lasR mutant CF416L1 relative to CF416 and CF416L1R. Values are the average cfu of two cultures, and error bars show standard deviations.
    Figure Legend Snippet: Inactivation of lasR confers increased β-lactamase activity and β-lactam tolerance. A. Ceftazidime-resistant colonies emerged in a lawn of CF416 or CF416L1 cells on LB agar with 20 µg ml −1 of ceftazidime (Petri dish upper and lower halves), but only the lasR mutant lawns yielded small partially resistant colonies whose growth is inhibited by the addition of 8 µg ml −1 of the β-lactamase inhibitor tazobactam (Petri dish lower halves). Photographs were taken after incubation overnight at 37°C followed by 3 days at 22°C. B. β-Lactamase activity in culture supernatants, measured as a change in optical density at 490 nm (ΔA 490 ) after addition of the chromogenic substrate nitrocefin. Cases with no change are marked with an asterisk. Values are the average of three technical replicates, and error bars show standard deviations. Equivalent results were obtained using a qualitative whole-culture assay. C. β-Lactamase activity in whole cultures of CF416 and CF416 lasR ::Gm. Values are the averages of three cultures, and error bars show standard error of the mean. One unit of β-lactamase is defined as 1 µmol of nitrocefin hydrolysed per min per mg of total protein. D. Reduced killing by ceftazidime of the lasR mutant CF416L1 relative to CF416 and CF416L1R. Values are the average cfu of two cultures, and error bars show standard deviations.

    Techniques Used: Activity Assay, Mutagenesis, Incubation

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    Activity Assay:

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    Article Snippet: .. d -Aminopeptidase activity was detected by monitoring the formation of p -nitroaniline from Gly-, d -Ala-, l -Ala-, and l -Ala- l -Ala- p -nitroanilide (Bachem) at 405 nm in a 100 m m Tris, 75 m m NaCl buffer (pH 7.5) at 25 °C. β-Lactamase activity was tested by monitoring the hydrolysis of nitrocefin (Calbiochem) at 482 nm in a 100 m m Tris, 75 m m NaCl buffer (pH 7.5) at 25 °C. .. ClbPpep (20 μ m ) was incubated at 20 °C for 24 h in a 100 m m Tris, 75 m m NaCl buffer (pH 7.5) alone, or with 20 m m imipenem (Sigma), in a 1-ml final volume.

    Article Title: Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria
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    Polymerase Chain Reaction:

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus
    Article Snippet: .. Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine. .. The type and amount of β-lactamase produced by each isolate following induction by growth on agar containing 0.5 μg of methicillin per ml were determined by using whole-cell suspensions of bacteria, as described previously ( ).

    Sequencing:

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus
    Article Snippet: .. Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine. .. The type and amount of β-lactamase produced by each isolate following induction by growth on agar containing 0.5 μg of methicillin per ml were determined by using whole-cell suspensions of bacteria, as described previously ( ).

    Synthesized:

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus
    Article Snippet: .. Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine. .. The type and amount of β-lactamase produced by each isolate following induction by growth on agar containing 0.5 μg of methicillin per ml were determined by using whole-cell suspensions of bacteria, as described previously ( ).

    Nucleic Acid Electrophoresis:

    Article Title: The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone
    Article Snippet: Cultures were separated into cell-bound and supernatant fractions by centrifugation, and total protein present in the supernatant fraction was precipitated with 12.5% trichloroacetic acid. .. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) by standard procedures , transferred to polyvinylidene difluoride membranes, and immunoblotted with anti-HA (Roche Diagnostics Corp., Indianapolis, Ind.), anti-NPTII (Cortex Biochem, San Leandro, Calif.), or anti-β-lactamase (Chemicon International, Temecula, Calif.) primary antibodies. .. Anti-β-lactamase and anti-NPTII primary antibodies were recognized with goat anti-rabbit immunoglobulin G-alkaline phosphatase conjugate (Sigma Chemical Co., St. Louis, Mo.), and anti-HA primary antibodies were recognized with goat anti-rat immunoglobulin G-alkaline phosphatase conjugate (Sigma Chemical Co.).

    SDS Page:

    Article Title: The Pseudomonas syringae HopPtoV Protein Is Secreted in Culture and Translocated into Plant Cells via the Type III Protein Secretion System in a Manner Dependent on the ShcV Type III Chaperone
    Article Snippet: Cultures were separated into cell-bound and supernatant fractions by centrifugation, and total protein present in the supernatant fraction was precipitated with 12.5% trichloroacetic acid. .. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) by standard procedures , transferred to polyvinylidene difluoride membranes, and immunoblotted with anti-HA (Roche Diagnostics Corp., Indianapolis, Ind.), anti-NPTII (Cortex Biochem, San Leandro, Calif.), or anti-β-lactamase (Chemicon International, Temecula, Calif.) primary antibodies. .. Anti-β-lactamase and anti-NPTII primary antibodies were recognized with goat anti-rabbit immunoglobulin G-alkaline phosphatase conjugate (Sigma Chemical Co., St. Louis, Mo.), and anti-HA primary antibodies were recognized with goat anti-rat immunoglobulin G-alkaline phosphatase conjugate (Sigma Chemical Co.).

    Incubation:

    Article Title: Copper Influences the Antibacterial Outcomes of a β-Lactamase-Activated Prochelator against Drug-Resistant Bacteria
    Article Snippet: LC-MS was run and peaks corresponding to PcephPT (11 min) and internal standard (14 min) were integrated in the UV chromatogram at 280 nm. .. Samples were prepared in 100 μL phosphate-buffered saline, pH 7.4 (Lonza) with 100 μM PcephPT, 10 μM Rofecoxib (as internal standard) and either 0.2 U/mL β-lactamase from E. cloacae (Type III, Sigma Aldrich) or no enzyme and incubated in closed tubes at r.t. Aliquots (20 μL) were taken at the indicated timepoints and immediately added to new tubes containing β-lactamase inhibitor tazobactam (2.5 mM in 5 μL water). .. These tubes were immediately mixed, sonicated, and frozen until shortly before LC-MS injection.

    Article Title: Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein
    Article Snippet: .. The β-lactamase gene was induced by adding isopropyl-β- d -thiogalactopyranoside (IPTG) to a final concentration of 0.5 mM and further incubation at 25°C for 4 h. Following induction the cells were pelleted and the supernatant containing the secreted, soluble β-lactamase was concentrated to 100 ml with an Amicon Centriprep-10 concentrator (Millipore Corp.). ..

    Isolation:

    Article Title: Synergism and the mechanism of action of the combination of α-mangostin isolated from Garcinia mangostana L. and oxacillin against an oxacillin-resistant Staphylococcus saprophyticus
    Article Snippet: The susceptible strain S. aureus ATCC 29213, a reference strain, was obtained from the American Type Culture Collection (ATCC). .. Oxacillin, Nisin, o-nitrophenol-β-D-galactoside (ONPG), α-mangostin standard, and β-lactamase type IV isolated from E. cloacae were obtained from Sigma-Aldrich, UK. .. Meuller-Hinton broth (MHB) and Mueller-Hinton agar (MHA) were purchased from Oxoid (Basingstoke, UK).

    other:

    Article Title: The Synergy and Mode of Action of Cyperus rotundus L. Extract Plus Ampicillin against Ampicillin-Resistant Staphylococcus aureus
    Article Snippet: Chemicals and Media Ampicillin, nisin, and β -lactamase type IV from Enterobacter cloacae were obtained from Sigma-Aldrich.

    Concentration Assay:

    Article Title: Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein
    Article Snippet: .. The β-lactamase gene was induced by adding isopropyl-β- d -thiogalactopyranoside (IPTG) to a final concentration of 0.5 mM and further incubation at 25°C for 4 h. Following induction the cells were pelleted and the supernatant containing the secreted, soluble β-lactamase was concentrated to 100 ml with an Amicon Centriprep-10 concentrator (Millipore Corp.). ..

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  • 94
    Millipore β lactamase type iv
    The inhibitory activity of SSE against <t>β</t> -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p
    β Lactamase Type Iv, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore β lactamase activity
    Effect of cheopin on Y. pestis outer membranes. The extent of OM damage caused by cheopin was determined by measuring the release of <t>β-lactamase</t> with the colorimetric substrate CENTA. Y. pestis strains expressing β-lactamase were treated with 4μM cheopin or FastBreak™ cell lysis solution (positive control) and the hydrolysis of CENTA was monitored for 1h at 28 °C. A: Y. pestis KIM6+ (pUC19), B: Y. pestis KIM6+ΔarnOP (pUC19), C: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77) and D: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77-arnOP).
    β Lactamase Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore penicillinase
    Sterigmatocystin (A) and penicillin (B) production in xprG loss-of-function ( xprG - ) and gain-of-function ( xprG1 ) mutants. A. Sterigmatocystin, extracted from the filtered growth medium of an xprG + strain (MH2), an xprG1 strain (MK85) and two xprG - strains, MK198 ( xprG2) and MK422 ( xprGΔ1 ), was analyzed using thin layer chromatography. Sterigmatocystin fluoresces yellow after treatment with AlCl 3 . Sterigmatocystin (ST) (Sigma) was applied as a standard. The cultures used in the assays were generated by inoculating growth medium with 3 x 10 8 conidia. After transfer to carbon-free medium for 24 h, the dry mycelial weights were 100 mg ( xprG + ), 71 mg ( xprG1 ), 129 ( xprG2 ) and 152 mg ( xprGΔ1 ). B. Penicillin bioassay based on inhibition of bacterial growth. Samples of filtered, concentrated growth medium from strains MH2 ( xprG + ), MK85 ( xprG1 ), and MK198 ( xprG2 ) was applied to wells in medium seeded with the Micrococcus luteus . 400 ng of penicillin G (penG) and 10 mM sodium orthophosphate buffer pH 6.8 (buffer) were used as controls. The Aspergillus growth medium contained either 3% glucose or 3% lactose. In the right-hand plate the samples were treated with 1 U of <t>penicillinase</t> (Sigma Aldrich) before they were applied to the wells. The full genotypes of the strains are given in Table 1 .
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    beta Lactamase Testkit is a rapid acidimetric test for detection of beta lactamase activity of microorganisms The test is performed in microtitre wells This test is based on hydrolysis of
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    The inhibitory activity of SSE against β -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: The Synergy and Mode of Action of Cyperus rotundus L. Extract Plus Ampicillin against Ampicillin-Resistant Staphylococcus aureus

    doi: 10.1155/2018/3438453

    Figure Lengend Snippet: The inhibitory activity of SSE against β -lactamase type IV from E. cloacae in hydrolyzing benzylpenicillin; CON = control (no testing agent); AMP (32) = ampicillin at 32 μ g/ml; CRE (0.25) = CRE at 0.25 mg/ml; AMP (0.5) + CRE (0.0625) = ampicillin at 0.5 μ g/ml plus CRE at 0.0625 mg/ml. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Tukey's HSD, p

    Article Snippet: Chemicals and Media Ampicillin, nisin, and β -lactamase type IV from Enterobacter cloacae were obtained from Sigma-Aldrich.

    Techniques: Activity Assay

    Agarose gel (a) and Southern hybridization with a blaZ probe (b) of Eco RI-restricted plasmid DNA from phage group II, β-lactamase-producing isolates of S. aureus ).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus

    doi:

    Figure Lengend Snippet: Agarose gel (a) and Southern hybridization with a blaZ probe (b) of Eco RI-restricted plasmid DNA from phage group II, β-lactamase-producing isolates of S. aureus ).

    Article Snippet: Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine.

    Techniques: Agarose Gel Electrophoresis, Hybridization, Plasmid Preparation

    Southern hybridization of chromosomal DNA from phage group II, type B β-lactamase-producing isolates of S. aureus using a blaZ probe. Lanes 1 to 9, Eco RI-restricted DNA; lanes 10 to 18, Hin ).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus

    doi:

    Figure Lengend Snippet: Southern hybridization of chromosomal DNA from phage group II, type B β-lactamase-producing isolates of S. aureus using a blaZ probe. Lanes 1 to 9, Eco RI-restricted DNA; lanes 10 to 18, Hin ).

    Article Snippet: Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine.

    Techniques: Hybridization

    Comparison of the nucleotide sequence of the gene encoding type B β-lactamase from strain 22260 to other S. aureus ). Differences in nucleotide sequences are shown; −, no change from the blaZ ). L1 to L24 refer to the leader peptide. Whereas the sequences of the type A, C, and D β-lactamases have been determined through the carboxy-terminal residue at Ambler position 290, reliable sequence data for the type B enzyme were obtained through residue 253.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Characterization of a Chromosomal Gene Encoding Type B ?-Lactamase in Phage Group II Isolates of Staphylococcus aureus

    doi:

    Figure Lengend Snippet: Comparison of the nucleotide sequence of the gene encoding type B β-lactamase from strain 22260 to other S. aureus ). Differences in nucleotide sequences are shown; −, no change from the blaZ ). L1 to L24 refer to the leader peptide. Whereas the sequences of the type A, C, and D β-lactamases have been determined through the carboxy-terminal residue at Ambler position 290, reliable sequence data for the type B enzyme were obtained through residue 253.

    Article Snippet: Oligonucleotide primers for PCR and for sequencing of the type B β-lactamase gene were synthesized on a Cyclone Plus Automatic DNA synthesizer (Millipore, Bedford, Mass.) by the DNA Core Facility, Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine.

    Techniques: Sequencing

    Effect of cheopin on Y. pestis outer membranes. The extent of OM damage caused by cheopin was determined by measuring the release of β-lactamase with the colorimetric substrate CENTA. Y. pestis strains expressing β-lactamase were treated with 4μM cheopin or FastBreak™ cell lysis solution (positive control) and the hydrolysis of CENTA was monitored for 1h at 28 °C. A: Y. pestis KIM6+ (pUC19), B: Y. pestis KIM6+ΔarnOP (pUC19), C: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77) and D: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77-arnOP).

    Journal: bioRxiv

    Article Title: Yersinia pestis lipopolysaccharide remodeling confers resistance to a Xenopsylla cheopis cecropin

    doi: 10.1101/2021.03.12.435208

    Figure Lengend Snippet: Effect of cheopin on Y. pestis outer membranes. The extent of OM damage caused by cheopin was determined by measuring the release of β-lactamase with the colorimetric substrate CENTA. Y. pestis strains expressing β-lactamase were treated with 4μM cheopin or FastBreak™ cell lysis solution (positive control) and the hydrolysis of CENTA was monitored for 1h at 28 °C. A: Y. pestis KIM6+ (pUC19), B: Y. pestis KIM6+ΔarnOP (pUC19), C: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77) and D: Y. pestis KIM6+ ΔarnOP (pUC19, pMWO77-arnOP).

    Article Snippet: Outer Membrane Permeabilization Assay The effects of cheopin on the integrity of the bacterial outer membranes were studied by measuring the β-lactamase activity as previously described [ ] with slight modifications.

    Techniques: Expressing, Lysis, Positive Control

    Sterigmatocystin (A) and penicillin (B) production in xprG loss-of-function ( xprG - ) and gain-of-function ( xprG1 ) mutants. A. Sterigmatocystin, extracted from the filtered growth medium of an xprG + strain (MH2), an xprG1 strain (MK85) and two xprG - strains, MK198 ( xprG2) and MK422 ( xprGΔ1 ), was analyzed using thin layer chromatography. Sterigmatocystin fluoresces yellow after treatment with AlCl 3 . Sterigmatocystin (ST) (Sigma) was applied as a standard. The cultures used in the assays were generated by inoculating growth medium with 3 x 10 8 conidia. After transfer to carbon-free medium for 24 h, the dry mycelial weights were 100 mg ( xprG + ), 71 mg ( xprG1 ), 129 ( xprG2 ) and 152 mg ( xprGΔ1 ). B. Penicillin bioassay based on inhibition of bacterial growth. Samples of filtered, concentrated growth medium from strains MH2 ( xprG + ), MK85 ( xprG1 ), and MK198 ( xprG2 ) was applied to wells in medium seeded with the Micrococcus luteus . 400 ng of penicillin G (penG) and 10 mM sodium orthophosphate buffer pH 6.8 (buffer) were used as controls. The Aspergillus growth medium contained either 3% glucose or 3% lactose. In the right-hand plate the samples were treated with 1 U of penicillinase (Sigma Aldrich) before they were applied to the wells. The full genotypes of the strains are given in Table 1 .

    Journal: F1000Research

    Article Title: A p53-like transcription factor similar to Ndt80 controls the response to nutrient stress in the filamentous fungus,Aspergillus nidulans

    doi: 10.12688/f1000research.2-72.v1

    Figure Lengend Snippet: Sterigmatocystin (A) and penicillin (B) production in xprG loss-of-function ( xprG - ) and gain-of-function ( xprG1 ) mutants. A. Sterigmatocystin, extracted from the filtered growth medium of an xprG + strain (MH2), an xprG1 strain (MK85) and two xprG - strains, MK198 ( xprG2) and MK422 ( xprGΔ1 ), was analyzed using thin layer chromatography. Sterigmatocystin fluoresces yellow after treatment with AlCl 3 . Sterigmatocystin (ST) (Sigma) was applied as a standard. The cultures used in the assays were generated by inoculating growth medium with 3 x 10 8 conidia. After transfer to carbon-free medium for 24 h, the dry mycelial weights were 100 mg ( xprG + ), 71 mg ( xprG1 ), 129 ( xprG2 ) and 152 mg ( xprGΔ1 ). B. Penicillin bioassay based on inhibition of bacterial growth. Samples of filtered, concentrated growth medium from strains MH2 ( xprG + ), MK85 ( xprG1 ), and MK198 ( xprG2 ) was applied to wells in medium seeded with the Micrococcus luteus . 400 ng of penicillin G (penG) and 10 mM sodium orthophosphate buffer pH 6.8 (buffer) were used as controls. The Aspergillus growth medium contained either 3% glucose or 3% lactose. In the right-hand plate the samples were treated with 1 U of penicillinase (Sigma Aldrich) before they were applied to the wells. The full genotypes of the strains are given in Table 1 .

    Article Snippet: The filtrates were left to diffuse for 18 h at 4°C and then incubated at 30°C for 32 h. For samples treated with penicillinase (Sigma Aldrich), 1 µL containing 1 U of enzyme in 100 mM Tris-HCl pH7 with 0.1% BSA was added and the samples were incubated at 25°C for 15 min before they were applied to the plates.

    Techniques: Thin Layer Chromatography, Generated, Inhibition