bl21 bacterial cells  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 bacterial cells
    Bl21 Bacterial Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 bacterial cells/product/Thermo Fisher
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bl21 bacterial cells - by Bioz Stars, 2020-09
    88/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Identification of Osteopontin Phosphorylation Sites Involved in Bone Remodeling and Inhibition of Pathological Calcification
    Article Snippet: .. Clones displaying integrate cDNA sequence and correct reading frame were selected for protein expression in BL21 bacterial cells (Invitrogen) under protocols described elsewhere [ ]. .. Thrombin (Amersham) was mixed with aliquots of the beads-GST-fusion protein at a ratio of 2–4 U/mg in 1× PBS.

    In Vitro:

    Article Title: MDM2 Inhibits Axin-Induced p53 Activation Independently of its E3 Ligase Activity
    Article Snippet: .. In vitro Binding Assay The proteins His-Axin, His-p53, GST-MDM2, GST-MDM2 (C464A) and GST- MDM2Δp53 were expressed in BL21 bacterial cells (purchased from Invitrogen) induced by 1 mM IPTG for 6 h at 26°C, then were purified using His-select nickel affinity gel (Sigma) or glutathione-agarose beads (GE). ..

    Construct:

    Article Title: Hsp90 Directly Modulates the Spatial Distribution of AF9/MLLT3 and Affects Target Gene Expression *
    Article Snippet: .. This plasmid construct was transformed into BL21 bacterial cells (Invitrogen) for protein expression. .. Following induction of protein expression with isopropyl β- d -1-thiogalactopyranoside (IPTG), cell pellets were collected and resuspended in bacterial lysis buffer (20 m m Tris, pH 7.4, 1 m NaCl, 1 m m EDTA, and 1 m m dithiothreitol).

    Purification:

    Article Title: MDM2 Inhibits Axin-Induced p53 Activation Independently of its E3 Ligase Activity
    Article Snippet: .. In vitro Binding Assay The proteins His-Axin, His-p53, GST-MDM2, GST-MDM2 (C464A) and GST- MDM2Δp53 were expressed in BL21 bacterial cells (purchased from Invitrogen) induced by 1 mM IPTG for 6 h at 26°C, then were purified using His-select nickel affinity gel (Sigma) or glutathione-agarose beads (GE). ..

    Article Title: Regulation of Adipocyte Differentiation by the Zinc Finger Protein ZNF638 *
    Article Snippet: .. GST Pulldown Assays GST-ZNF638 fragments were purified from BL21 bacterial cells (Invitrogen) according to the following protocol. .. Cells were collected, pelleted, and lysed in 2 ml of lysis buffer (20 mm Tris (pH 8), 400 mm KCl, 1 mm EDTA, 0.1% Triton X-100, and 1 mm DTT).

    Expressing:

    Article Title: Hsp90 Directly Modulates the Spatial Distribution of AF9/MLLT3 and Affects Target Gene Expression *
    Article Snippet: .. This plasmid construct was transformed into BL21 bacterial cells (Invitrogen) for protein expression. .. Following induction of protein expression with isopropyl β- d -1-thiogalactopyranoside (IPTG), cell pellets were collected and resuspended in bacterial lysis buffer (20 m m Tris, pH 7.4, 1 m NaCl, 1 m m EDTA, and 1 m m dithiothreitol).

    Article Title: Identification of Osteopontin Phosphorylation Sites Involved in Bone Remodeling and Inhibition of Pathological Calcification
    Article Snippet: .. Clones displaying integrate cDNA sequence and correct reading frame were selected for protein expression in BL21 bacterial cells (Invitrogen) under protocols described elsewhere [ ]. .. Thrombin (Amersham) was mixed with aliquots of the beads-GST-fusion protein at a ratio of 2–4 U/mg in 1× PBS.

    Sequencing:

    Article Title: Identification of Osteopontin Phosphorylation Sites Involved in Bone Remodeling and Inhibition of Pathological Calcification
    Article Snippet: .. Clones displaying integrate cDNA sequence and correct reading frame were selected for protein expression in BL21 bacterial cells (Invitrogen) under protocols described elsewhere [ ]. .. Thrombin (Amersham) was mixed with aliquots of the beads-GST-fusion protein at a ratio of 2–4 U/mg in 1× PBS.

    Transformation Assay:

    Article Title: Hsp90 Directly Modulates the Spatial Distribution of AF9/MLLT3 and Affects Target Gene Expression *
    Article Snippet: .. This plasmid construct was transformed into BL21 bacterial cells (Invitrogen) for protein expression. .. Following induction of protein expression with isopropyl β- d -1-thiogalactopyranoside (IPTG), cell pellets were collected and resuspended in bacterial lysis buffer (20 m m Tris, pH 7.4, 1 m NaCl, 1 m m EDTA, and 1 m m dithiothreitol).

    Binding Assay:

    Article Title: MDM2 Inhibits Axin-Induced p53 Activation Independently of its E3 Ligase Activity
    Article Snippet: .. In vitro Binding Assay The proteins His-Axin, His-p53, GST-MDM2, GST-MDM2 (C464A) and GST- MDM2Δp53 were expressed in BL21 bacterial cells (purchased from Invitrogen) induced by 1 mM IPTG for 6 h at 26°C, then were purified using His-select nickel affinity gel (Sigma) or glutathione-agarose beads (GE). ..

    Plasmid Preparation:

    Article Title: Hsp90 Directly Modulates the Spatial Distribution of AF9/MLLT3 and Affects Target Gene Expression *
    Article Snippet: .. This plasmid construct was transformed into BL21 bacterial cells (Invitrogen) for protein expression. .. Following induction of protein expression with isopropyl β- d -1-thiogalactopyranoside (IPTG), cell pellets were collected and resuspended in bacterial lysis buffer (20 m m Tris, pH 7.4, 1 m NaCl, 1 m m EDTA, and 1 m m dithiothreitol).

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    Thermo Fisher e coli bl21
    Spot assays used for monitoring the growth performance of <t>BL21/pEZYHb</t> and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress
    E Coli Bl21, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21/product/Thermo Fisher
    Average 99 stars, based on 151 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher bl21 cells
    Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli <t>BL21</t> control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.
    Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 cells/product/Thermo Fisher
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    bl21 cells - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher e coli bl21 cells
    Spot assays used for monitoring the growth performance of <t>BL21/pEZYHb</t> and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress
    E Coli Bl21 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli bl21 cells/product/Thermo Fisher
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    e coli bl21 cells - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Journal: BMC Genomics

    Article Title: The alcohol dehydrogenase gene family in sugarcane and its involvement in cold stress regulation

    doi: 10.1186/s12864-020-06929-9

    Figure Lengend Snippet: Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Article Snippet: E. coli BL21 with the empty vector pEZYHb was used as the control.

    Techniques: Transformation Assay, Incubation

    The serine catalytic site is necessary for the cytopathic effect of TagB and TagC. ( A ) Concentrated supernatants containing 30 μg of protein per well derived from E. coli BL21 clones expressing TagB, TagC, Sha, or their respective serine mutant variant proteins were incubated with monolayers of the 5637 human bladder epithelial cell line for 5 h at 37 °C. Cytopathic effects (white triangle) were absent in cells treated with the serine catalytic site mutant variants of TagB or TagC. The empty vector (pBCsk+) without insert was used as a negative control. The scale bar represents 20 µm. ( B ) Cytotoxicity measured by LDH release from 5637 human bladder cells after incubation with supernatant filtrates of different clones (30 μg of protein per well) at 37 °C for 5 h. Empty vector (pBCsk+) was used as a negative control and maximum LDH release (positive control) was determined by treatment with lysis solution. Data are the means of three independent experiments, and error bars represent the standard errors of the means. Significant differences between lysis caused by native and mutant SPATEs were determined using Student’s t -test with *** p

    Journal: International Journal of Molecular Sciences

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    doi: 10.3390/ijms21093047

    Figure Lengend Snippet: The serine catalytic site is necessary for the cytopathic effect of TagB and TagC. ( A ) Concentrated supernatants containing 30 μg of protein per well derived from E. coli BL21 clones expressing TagB, TagC, Sha, or their respective serine mutant variant proteins were incubated with monolayers of the 5637 human bladder epithelial cell line for 5 h at 37 °C. Cytopathic effects (white triangle) were absent in cells treated with the serine catalytic site mutant variants of TagB or TagC. The empty vector (pBCsk+) without insert was used as a negative control. The scale bar represents 20 µm. ( B ) Cytotoxicity measured by LDH release from 5637 human bladder cells after incubation with supernatant filtrates of different clones (30 μg of protein per well) at 37 °C for 5 h. Empty vector (pBCsk+) was used as a negative control and maximum LDH release (positive control) was determined by treatment with lysis solution. Data are the means of three independent experiments, and error bars represent the standard errors of the means. Significant differences between lysis caused by native and mutant SPATEs were determined using Student’s t -test with *** p

    Article Snippet: Electron Microscopy Immunogold labeling of bacteria was carried out by culturing E. coli BL21 expressing different SPATEs in Luria-Bertani medium supplemented with 30 µg/mL chloramphenicol for 5 h. Bacterial suspensions (50 µL) were spotted on nickel-coated TEM grids.

    Techniques: Derivative Assay, Clone Assay, Expressing, Mutagenesis, Variant Assay, Incubation, Plasmid Preparation, Negative Control, Positive Control, Lysis

    The autoaggregation phenotype is independent of the serine protease motif. Clones of E. coli BL21 expressing TagB, TagC, Sha, or their respective serine-site mutants were grown 18 h and adjusted to an OD 600 of 1.5 and left to rest at 4 °C. Samples were taken at 1 cm from the top surface of the cultures after 3 h to determine the change in OD 600 . Assays were performed in triplicate, and the rate of autoaggregation was determined by the mean decrease in OD 600nm after 3 h. E. coli BL21 pBCsk+ vector without insert (empty vector) was used as a negative control and the AIDA-1 autotransporter was the positive control for autoaggregation. Error bars represent standard errors of the means (*** p

    Journal: International Journal of Molecular Sciences

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    doi: 10.3390/ijms21093047

    Figure Lengend Snippet: The autoaggregation phenotype is independent of the serine protease motif. Clones of E. coli BL21 expressing TagB, TagC, Sha, or their respective serine-site mutants were grown 18 h and adjusted to an OD 600 of 1.5 and left to rest at 4 °C. Samples were taken at 1 cm from the top surface of the cultures after 3 h to determine the change in OD 600 . Assays were performed in triplicate, and the rate of autoaggregation was determined by the mean decrease in OD 600nm after 3 h. E. coli BL21 pBCsk+ vector without insert (empty vector) was used as a negative control and the AIDA-1 autotransporter was the positive control for autoaggregation. Error bars represent standard errors of the means (*** p

    Article Snippet: Electron Microscopy Immunogold labeling of bacteria was carried out by culturing E. coli BL21 expressing different SPATEs in Luria-Bertani medium supplemented with 30 µg/mL chloramphenicol for 5 h. Bacterial suspensions (50 µL) were spotted on nickel-coated TEM grids.

    Techniques: Clone Assay, Expressing, Plasmid Preparation, Negative Control, Positive Control

    ( A ) Silver stained SDS-PAGE analysis of concentrated supernatants of E. coli BL21 expressing SPATE proteins. Filtered supernatants from clones expressing TagB, TagC, and Sha or the variant TagB S255A, TagC S252A, and Sha S258A proteins were concentrated through Amicon filters with a 50 kDa cutoff. Samples containing 5 µg of protein were migrated and stained with silver stain. ( B – E ) Immunogold Electron Microscopy (EM) of SPATEs (Serine protease autotransporters of Enterobacteriaceae ) localized to the outer membrane and extracellular medium. Immunogold-TEM micrographs of SPATEs using SPATE-specific antiserum. Bacteria were cultured to 0.6 OD 600nm in Luria-Bertani medium. E. coli BL21 pBCsk+ expressing TagB ( B ), TagC ( C ), and Sha ( D ) labelled with immunogold particles. ( E ) E. coli BL21 pBcsk+ (vector only control) shows no immunogold staining. Insets represents boxed areas of higher magnification showing clustering of SPATE proteins. All images were acquired at ×17,000 magnification; scale bars represent 1 µm, and 0.5 µm (Insets).

    Journal: International Journal of Molecular Sciences

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    doi: 10.3390/ijms21093047

    Figure Lengend Snippet: ( A ) Silver stained SDS-PAGE analysis of concentrated supernatants of E. coli BL21 expressing SPATE proteins. Filtered supernatants from clones expressing TagB, TagC, and Sha or the variant TagB S255A, TagC S252A, and Sha S258A proteins were concentrated through Amicon filters with a 50 kDa cutoff. Samples containing 5 µg of protein were migrated and stained with silver stain. ( B – E ) Immunogold Electron Microscopy (EM) of SPATEs (Serine protease autotransporters of Enterobacteriaceae ) localized to the outer membrane and extracellular medium. Immunogold-TEM micrographs of SPATEs using SPATE-specific antiserum. Bacteria were cultured to 0.6 OD 600nm in Luria-Bertani medium. E. coli BL21 pBCsk+ expressing TagB ( B ), TagC ( C ), and Sha ( D ) labelled with immunogold particles. ( E ) E. coli BL21 pBcsk+ (vector only control) shows no immunogold staining. Insets represents boxed areas of higher magnification showing clustering of SPATE proteins. All images were acquired at ×17,000 magnification; scale bars represent 1 µm, and 0.5 µm (Insets).

    Article Snippet: Electron Microscopy Immunogold labeling of bacteria was carried out by culturing E. coli BL21 expressing different SPATEs in Luria-Bertani medium supplemented with 30 µg/mL chloramphenicol for 5 h. Bacterial suspensions (50 µL) were spotted on nickel-coated TEM grids.

    Techniques: Staining, SDS Page, Expressing, Clone Assay, Variant Assay, Silver Staining, Electron Microscopy, Transmission Electron Microscopy, Cell Culture, Plasmid Preparation

    Effects of TagB, TagC, and Sha on the actin cytoskeleton of bladder epithelial cells is serine-protease-motif dependent. ( A ) Concentrated supernatant extracts (30 μg of protein per well) from E. coli BL21 clones expressing TagB, TagC, or Sha and their respective serine catalytic site mutants were incubated with monolayers of human bladder (5637) epithelial cells for 5 h at 37 °C. After incubation, cells were fixed and permeabilized. Actin was stained with fluorescently labeled phalloidin (green) and the nucleus was stained by DAPI (blue). Cells treated with the filtered supernatant of E. coli BL21 pBCsk+ without insert (empty vector) were used as a negative control. Slides were observed by confocal microscopy. Inset images from the left panels are magnified in the panels to the right. Bars represent 10 µm. ( B ) Quantitative analysis of fluorescence intensity of F-actin. Analysis of fluorescent intensity was done at the original magnification by measuring the mean gray value with ImageJ software [ 22 ] with an n value of at least 10 cells. Data values represent the mean ± SEM of at least three independent experiments. (* p

    Journal: International Journal of Molecular Sciences

    Article Title: The Serine Protease Autotransporters TagB, TagC, and Sha from Extraintestinal Pathogenic Escherichia coli Are Internalized by Human Bladder Epithelial Cells and Cause Actin Cytoskeletal Disruption

    doi: 10.3390/ijms21093047

    Figure Lengend Snippet: Effects of TagB, TagC, and Sha on the actin cytoskeleton of bladder epithelial cells is serine-protease-motif dependent. ( A ) Concentrated supernatant extracts (30 μg of protein per well) from E. coli BL21 clones expressing TagB, TagC, or Sha and their respective serine catalytic site mutants were incubated with monolayers of human bladder (5637) epithelial cells for 5 h at 37 °C. After incubation, cells were fixed and permeabilized. Actin was stained with fluorescently labeled phalloidin (green) and the nucleus was stained by DAPI (blue). Cells treated with the filtered supernatant of E. coli BL21 pBCsk+ without insert (empty vector) were used as a negative control. Slides were observed by confocal microscopy. Inset images from the left panels are magnified in the panels to the right. Bars represent 10 µm. ( B ) Quantitative analysis of fluorescence intensity of F-actin. Analysis of fluorescent intensity was done at the original magnification by measuring the mean gray value with ImageJ software [ 22 ] with an n value of at least 10 cells. Data values represent the mean ± SEM of at least three independent experiments. (* p

    Article Snippet: Electron Microscopy Immunogold labeling of bacteria was carried out by culturing E. coli BL21 expressing different SPATEs in Luria-Bertani medium supplemented with 30 µg/mL chloramphenicol for 5 h. Bacterial suspensions (50 µL) were spotted on nickel-coated TEM grids.

    Techniques: Clone Assay, Expressing, Incubation, Staining, Labeling, Plasmid Preparation, Negative Control, Confocal Microscopy, Fluorescence, Software

    Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.

    Journal: bioRxiv

    Article Title: The DNA mimic protein BCAS0292 is involved in the regulation of virulence of Burkholderia cenocepacia

    doi: 10.1101/2020.08.06.226779

    Figure Lengend Snippet: Role of BCAS0292 in epithelial cell attachment. A) Attachment of K56-2 WT, ΔBCAS0292 mutant and Δ0292_0292 complemented strain to CFBE41o − cells at an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain with 20 fields of view counted per data point in three independent experiments, ** p =0.0073. B) Confocal microscopy images representing the attachment of E. coli BL21 control and E. coli BL21_BCAS0292 strains to CFBE41o-cells. Bacteria were labelled using a primary anti- E. coli FITC-conjugated antibody (green). Nuclei of CFBE41o-cells were counterstained with DAPI (blue) and actin stained with phalloidin conjugated with Alexa fluor 568. C) Attachment E. coli BL21 control and E. coli BL21_BCAS0292 to CFBE41o-cells, using an MOI 50:1. Data represents the mean number of bacteria/ 100 cells for each strain, with 20 fields of view counted per data point in three independent experiments. D) Confirmation of expression of BCAS0292, following induction in E. coli BL21 cells with 1mM IPTG by Coomassie Blue stained SDS PAGE gel (conditions used for Figure 1C ). Lane 1: MW marker; Lane 2: E. coli BL21 expression control; Lane 3: E. coli BL21_BCAS0292 after incubation with IPTG (1mM) for 2 h.

    Article Snippet: Following the confirmation of recombinant BCAS0292 (rBCAS0292) expression in a pilot study, 1 L cultures of BL21 cells transformed with the expression plasmid, were grown in LB-ampicillin (100 μg/ ml) and induced with IPTG at a final concentration of 1 mM overnight at 25 °C.

    Techniques: Cell Attachment Assay, Mutagenesis, Confocal Microscopy, Staining, Expressing, SDS Page, Marker, Incubation

    Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Journal: BMC Genomics

    Article Title: The alcohol dehydrogenase gene family in sugarcane and its involvement in cold stress regulation

    doi: 10.1186/s12864-020-06929-9

    Figure Lengend Snippet: Spot assays used for monitoring the growth performance of BL21/pEZYHb and BL21/pEZYHb- ScADH3 transformed E. coli cells on LB plates under cold stress. The growth performance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells on LB plates without supplements or stresses were used as controls. After spotting the sample on LB agar plates, incubation at 4 °C in darkness for 3 days, 7 days, and 14 days was used to study the tolerance of BL21/pEZYHb and BL21/ pEZYHb- ScADH3 cells under cold stress

    Article Snippet: When E. coli BL21 cells containing different vectors were grown to OD600 of 0.6, 1.0 mmol L− 1 isopropyl β-D-thiogalactoside (IPTG) was added to induce protein production.

    Techniques: Transformation Assay, Incubation