Structured Review

Toyobo dh5α
Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into <t>DH5α.</t> ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.
Dh5α, supplied by Toyobo, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 1 article reviews
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92/100 stars

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1) Product Images from "An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease"

Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease

Journal: The EMBO Journal

doi: 10.1038/sj.emboj.7600049

Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into DH5α. ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.
Figure Legend Snippet: Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into DH5α. ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.

Techniques Used: Selection, Isolation, TUNEL Assay, Staining, Activation Assay, Activity Assay

Related Articles

Clone Assay:

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Article Title: Genome of ‘Ca. Desulfovibrio trichonymphae', an H2-oxidizing bacterium in a tripartite symbiotic system within a protist cell in the termite gut
Article Snippet: .. Amplification products were cloned using the Zero Blunt TOPO PCR Cloning Kit for Sequencing (Invitrogen) and Competent Quick DH5α (TOYOBO, Tokyo, Japan), and were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosciences, Foster City, CA, USA) on an ABI 3730xl DNA Analyzer, as described previously ( ). .. Phylogenetic analysis and codon adaptation index (CAI) calculation Maximum-likelihood trees were constructed using MEGA6 ( ).

Transfection:

Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease
Article Snippet: .. For the first screening, transfected cells were exposed to Tm for 24 h. For the second screening, cells were transfected with candidate plasmids from the first screening and collected after incubation with Tm for 48 h. Expression vectors harboring Rzs were isolated from surviving cells and used to transform DH5α (TOYOBO, Tokyo, Japan). .. Lines of SK-N-SH cells that had been semistably transfected with Rz expression vectors were treated with Tm (2 μg/ml) for 24 h. Apoptotic cells were stained by TUNEL or propidium iodide.

Amplification:

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Article Title: Genome of ‘Ca. Desulfovibrio trichonymphae', an H2-oxidizing bacterium in a tripartite symbiotic system within a protist cell in the termite gut
Article Snippet: .. Amplification products were cloned using the Zero Blunt TOPO PCR Cloning Kit for Sequencing (Invitrogen) and Competent Quick DH5α (TOYOBO, Tokyo, Japan), and were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosciences, Foster City, CA, USA) on an ABI 3730xl DNA Analyzer, as described previously ( ). .. Phylogenetic analysis and codon adaptation index (CAI) calculation Maximum-likelihood trees were constructed using MEGA6 ( ).

Agarose Gel Electrophoresis:

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Ligation:

Article Title: piggyBac Transposon-mediated Long-term Gene Expression in Mice
Article Snippet: .. The ligation products were used to transform E. coli Competent Quick DH5α (Toyobo) or E. coli HST08 Premium Competent Cells (Takara Bio). pDNA was isolated and purified using QIAprep Spin Miniprep Kit. .. Nucleotide sequences of the pDNA were sequenced by Fasmac sequencing service (Fasmac, Atsugi, Japan).

Isolation:

Article Title: piggyBac Transposon-mediated Long-term Gene Expression in Mice
Article Snippet: .. The ligation products were used to transform E. coli Competent Quick DH5α (Toyobo) or E. coli HST08 Premium Competent Cells (Takara Bio). pDNA was isolated and purified using QIAprep Spin Miniprep Kit. .. Nucleotide sequences of the pDNA were sequenced by Fasmac sequencing service (Fasmac, Atsugi, Japan).

Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease
Article Snippet: .. For the first screening, transfected cells were exposed to Tm for 24 h. For the second screening, cells were transfected with candidate plasmids from the first screening and collected after incubation with Tm for 48 h. Expression vectors harboring Rzs were isolated from surviving cells and used to transform DH5α (TOYOBO, Tokyo, Japan). .. Lines of SK-N-SH cells that had been semistably transfected with Rz expression vectors were treated with Tm (2 μg/ml) for 24 h. Apoptotic cells were stained by TUNEL or propidium iodide.

Purification:

Article Title: piggyBac Transposon-mediated Long-term Gene Expression in Mice
Article Snippet: .. The ligation products were used to transform E. coli Competent Quick DH5α (Toyobo) or E. coli HST08 Premium Competent Cells (Takara Bio). pDNA was isolated and purified using QIAprep Spin Miniprep Kit. .. Nucleotide sequences of the pDNA were sequenced by Fasmac sequencing service (Fasmac, Atsugi, Japan).

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Polymerase Chain Reaction:

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Article Title: Genome of ‘Ca. Desulfovibrio trichonymphae', an H2-oxidizing bacterium in a tripartite symbiotic system within a protist cell in the termite gut
Article Snippet: .. Amplification products were cloned using the Zero Blunt TOPO PCR Cloning Kit for Sequencing (Invitrogen) and Competent Quick DH5α (TOYOBO, Tokyo, Japan), and were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosciences, Foster City, CA, USA) on an ABI 3730xl DNA Analyzer, as described previously ( ). .. Phylogenetic analysis and codon adaptation index (CAI) calculation Maximum-likelihood trees were constructed using MEGA6 ( ).

Incubation:

Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease
Article Snippet: .. For the first screening, transfected cells were exposed to Tm for 24 h. For the second screening, cells were transfected with candidate plasmids from the first screening and collected after incubation with Tm for 48 h. Expression vectors harboring Rzs were isolated from surviving cells and used to transform DH5α (TOYOBO, Tokyo, Japan). .. Lines of SK-N-SH cells that had been semistably transfected with Rz expression vectors were treated with Tm (2 μg/ml) for 24 h. Apoptotic cells were stained by TUNEL or propidium iodide.

Expressing:

Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease
Article Snippet: .. For the first screening, transfected cells were exposed to Tm for 24 h. For the second screening, cells were transfected with candidate plasmids from the first screening and collected after incubation with Tm for 48 h. Expression vectors harboring Rzs were isolated from surviving cells and used to transform DH5α (TOYOBO, Tokyo, Japan). .. Lines of SK-N-SH cells that had been semistably transfected with Rz expression vectors were treated with Tm (2 μg/ml) for 24 h. Apoptotic cells were stained by TUNEL or propidium iodide.

Sequencing:

Article Title: Host-Symbiont Cospeciation of Termite-Gut Cellulolytic Protists of the Genera Teranympha and Eucomonympha and their Treponema Endosymbionts
Article Snippet: .. The amplified products were separated by agarose gel electrophoresis, purified, and then cloned using the Zero Blunt pCR4-TOPO PCR cloning kit for sequencing (Invitrogen, Carlsbad, CA, USA) and Competent Quick DH5α (Toyobo). .. Clones containing inserts of the expected size were picked and partially sequenced, and the complete DNA sequence of each representative clone was obtained.

Article Title: Genome of ‘Ca. Desulfovibrio trichonymphae', an H2-oxidizing bacterium in a tripartite symbiotic system within a protist cell in the termite gut
Article Snippet: .. Amplification products were cloned using the Zero Blunt TOPO PCR Cloning Kit for Sequencing (Invitrogen) and Competent Quick DH5α (TOYOBO, Tokyo, Japan), and were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosciences, Foster City, CA, USA) on an ABI 3730xl DNA Analyzer, as described previously ( ). .. Phylogenetic analysis and codon adaptation index (CAI) calculation Maximum-likelihood trees were constructed using MEGA6 ( ).

Recombinant:

Article Title: Rapid preparation of mutated influenza hemagglutinins for influenza virus pandemic prevention
Article Snippet: .. Microbial strains and culture media The DH5α (F− , end A1, hsd R17[rk − /mK + ], sup E44, thi -1, λ − , deo R, rec A1, gyr A96, pho A, φ80dlac ZΔM15, Δ[lac ZYA-arg F]U169) Escherichia coli strain (Toyobo, Osaka, Japan) was used as the host for recombinant DNA manipulation. ..

Plasmid Preparation:

Article Title: Transmembrane Topologies of Ca2+-permeable Mechanosensitive Channels MCA1 and MCA2 in Arabidopsis thaliana *
Article Snippet: .. Strain XL1-Blue ( supE 44 hsdR 17 recA 1 endA 1 gyrA 46 thi relA 1 lac − F' [ proAB + lacI q lacZ ΔM15 Tn 10 ( tet r )]; Agilent Technologies, Santa Clara, CA) and DH5α ( supE 44 Δ lac U169 (φ80 lacZ ΔM15) hsdR 17 recA 1 endA 1 gyrA 96 thi -1 relA 1; Toyobo Co., Tokyo, Japan) were used for plasmid construction. ..

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  • 92
    Toyobo dh5α
    Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into <t>DH5α.</t> ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.
    Dh5α, supplied by Toyobo, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dh5α/product/Toyobo
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dh5α - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    91
    Toyobo coli dh5α strain
    Fly survival experiments. Virulence of the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), E . coli <t>DH5α</t> (as a negative control), and WT in the presence of L -serine (10, 20, 30, 40, and 50 mM) was evaluated. Significant differences, based on the log-rank test, were observed between WT and DH5α (*: P
    Coli Dh5α Strain, supplied by Toyobo, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/coli dh5α strain/product/Toyobo
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    coli dh5α strain - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    91
    Toyobo escherichia coli dh5α cells
    Fly survival experiments. Virulence of the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), E . coli <t>DH5α</t> (as a negative control), and WT in the presence of L -serine (10, 20, 30, 40, and 50 mM) was evaluated. Significant differences, based on the log-rank test, were observed between WT and DH5α (*: P
    Escherichia Coli Dh5α Cells, supplied by Toyobo, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli dh5α cells/product/Toyobo
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    escherichia coli dh5α cells - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    89
    Toyobo escherichia coli dh5α strain
    Fly survival experiments. Virulence of the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), E . coli <t>DH5α</t> (as a negative control), and WT in the presence of L -serine (10, 20, 30, 40, and 50 mM) was evaluated. Significant differences, based on the log-rank test, were observed between WT and DH5α (*: P
    Escherichia Coli Dh5α Strain, supplied by Toyobo, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli dh5α strain/product/Toyobo
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    escherichia coli dh5α strain - by Bioz Stars, 2020-08
    89/100 stars
      Buy from Supplier

    Image Search Results


    Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into DH5α. ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.

    Journal: The EMBO Journal

    Article Title: An RNA-dependent protein kinase is involved in tunicamycin-induced apoptosis and Alzheimer's disease

    doi: 10.1038/sj.emboj.7600049

    Figure Lengend Snippet: Selection of Rzs that suppress Tm-induced cell death from a randomized library. ( A ) Schematic representation of the selection system. ( B ) Number of transformants after each round of selection. Vectors isolated from surviving cells were introduced into DH5α. ( C ) Individual Rzs, selected from the randomized library, were examined for their ability to protect cells against Tm-mediated apoptosis. SK-N-SH cells that expressed selected Rzs were exposed to 2 μg/ml Tm for 24 h. Apoptosis was assessed by TUNEL staining. ( D ) The activation of caspase-3 depended on the activation of PKR. Cells were treated with Tm for 24 h or untreated (control), and then caspase-3 activity was measured. Cells treated with z-VAD-fmk were examined as controls.

    Article Snippet: For the first screening, transfected cells were exposed to Tm for 24 h. For the second screening, cells were transfected with candidate plasmids from the first screening and collected after incubation with Tm for 48 h. Expression vectors harboring Rzs were isolated from surviving cells and used to transform DH5α (TOYOBO, Tokyo, Japan).

    Techniques: Selection, Isolation, TUNEL Assay, Staining, Activation Assay, Activity Assay

    Fly survival experiments. Virulence of the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), E . coli DH5α (as a negative control), and WT in the presence of L -serine (10, 20, 30, 40, and 50 mM) was evaluated. Significant differences, based on the log-rank test, were observed between WT and DH5α (*: P

    Journal: PLoS ONE

    Article Title: Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

    doi: 10.1371/journal.pone.0169367

    Figure Lengend Snippet: Fly survival experiments. Virulence of the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), E . coli DH5α (as a negative control), and WT in the presence of L -serine (10, 20, 30, 40, and 50 mM) was evaluated. Significant differences, based on the log-rank test, were observed between WT and DH5α (*: P

    Article Snippet: E . coli DH5α strain was used as a control strain that does not penetrate epithelial cell monolayers [ ] and was purchased from TOYOBO, Japan.

    Techniques: Mutagenesis, Negative Control

    Bacterial adherence to Caco-2 cells for the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), PAO1Tn:: flgE mutant ( ΔflgE ), E . coli DH5α (as a negative control), and WT in the presence of L -serine (20, 30, 40, and 50 mM). Bacterial adherence was determined based on the number of adhered bacteria per Caco-2 cell. The assay was performed in six replicates, and the results are expressed as the mean ± SD. Significant differences were observed between WT and ΔserA (*: P

    Journal: PLoS ONE

    Article Title: Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

    doi: 10.1371/journal.pone.0169367

    Figure Lengend Snippet: Bacterial adherence to Caco-2 cells for the wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), PAO1Tn:: flgE mutant ( ΔflgE ), E . coli DH5α (as a negative control), and WT in the presence of L -serine (20, 30, 40, and 50 mM). Bacterial adherence was determined based on the number of adhered bacteria per Caco-2 cell. The assay was performed in six replicates, and the results are expressed as the mean ± SD. Significant differences were observed between WT and ΔserA (*: P

    Article Snippet: E . coli DH5α strain was used as a control strain that does not penetrate epithelial cell monolayers [ ] and was purchased from TOYOBO, Japan.

    Techniques: Mutagenesis, Negative Control

    Penetration activity of P . aeruginosa strains. (A) The wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), and PAO1Tn:: flgE mutant ( ΔflgE ) were inoculated onto the apical surfaces of Caco-2 cell monolayers at an MOI of 100, and the number of bacteria in the basolateral medium was counted at 6 h after infection. The assay was performed in triplicate, and the results are expressed as the mean ± SD. E . coli DH5α was used as a negative control. *#: P

    Journal: PLoS ONE

    Article Title: Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

    doi: 10.1371/journal.pone.0169367

    Figure Lengend Snippet: Penetration activity of P . aeruginosa strains. (A) The wild-type strain (WT), PAO1Tn:: serA mutant ( ΔserA ), PAO1Tn:: serA (pUCP19- serA ) complementary strain (+ serA ), and PAO1Tn:: flgE mutant ( ΔflgE ) were inoculated onto the apical surfaces of Caco-2 cell monolayers at an MOI of 100, and the number of bacteria in the basolateral medium was counted at 6 h after infection. The assay was performed in triplicate, and the results are expressed as the mean ± SD. E . coli DH5α was used as a negative control. *#: P

    Article Snippet: E . coli DH5α strain was used as a control strain that does not penetrate epithelial cell monolayers [ ] and was purchased from TOYOBO, Japan.

    Techniques: Activity Assay, Mutagenesis, Infection, Negative Control

    PGDH activity assay. (A) SDS-PAGE analysis to verify overexpression of the P . aeruginosa serA and synthesis of SerA proteins in crude extract isolated from DH5α (ptac-85- serA ) culture (SerA) and from DH5α (ptac-85) culture (mock). The red arrow indicates an overexpressed band at the expected size of 44 kDa for P . aeruginosa SerA. (B) Inhibitory effect of 50 mM L -serine on PGDH activity of SerA proteins (mU/mg) in crude extract isolated from DH5α (ptac-85- serA ) culture (SerA) and from DH5α (ptac-85) culture (mock). The assay was performed in five replicates, and the results are expressed as the mean ± SD. Significant differences were observed between SerA and mock (*: P

    Journal: PLoS ONE

    Article Title: Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

    doi: 10.1371/journal.pone.0169367

    Figure Lengend Snippet: PGDH activity assay. (A) SDS-PAGE analysis to verify overexpression of the P . aeruginosa serA and synthesis of SerA proteins in crude extract isolated from DH5α (ptac-85- serA ) culture (SerA) and from DH5α (ptac-85) culture (mock). The red arrow indicates an overexpressed band at the expected size of 44 kDa for P . aeruginosa SerA. (B) Inhibitory effect of 50 mM L -serine on PGDH activity of SerA proteins (mU/mg) in crude extract isolated from DH5α (ptac-85- serA ) culture (SerA) and from DH5α (ptac-85) culture (mock). The assay was performed in five replicates, and the results are expressed as the mean ± SD. Significant differences were observed between SerA and mock (*: P

    Article Snippet: E . coli DH5α strain was used as a control strain that does not penetrate epithelial cell monolayers [ ] and was purchased from TOYOBO, Japan.

    Techniques: Activity Assay, SDS Page, Over Expression, Isolation

    Motility assays. (A) Swimming motility of WT, ΔserA , + serA , and E . coli DH5α (as a negative control). A representative image from the swimming motility assay is shown. The major axis of swimming is the longest length of the swimming area. The assay was performed in triplicate, and the results are expressed as the mean ± SD. Significant differences were observed between WT and ΔserA (*: P

    Journal: PLoS ONE

    Article Title: Pseudomonas aeruginosa serA Gene Is Required for Bacterial Translocation through Caco-2 Cell Monolayers

    doi: 10.1371/journal.pone.0169367

    Figure Lengend Snippet: Motility assays. (A) Swimming motility of WT, ΔserA , + serA , and E . coli DH5α (as a negative control). A representative image from the swimming motility assay is shown. The major axis of swimming is the longest length of the swimming area. The assay was performed in triplicate, and the results are expressed as the mean ± SD. Significant differences were observed between WT and ΔserA (*: P

    Article Snippet: E . coli DH5α strain was used as a control strain that does not penetrate epithelial cell monolayers [ ] and was purchased from TOYOBO, Japan.

    Techniques: Negative Control, Motility Assay