bl21 star de3  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 star de3
    Systematic study of AND gate circuit overlap domain lengths and comparison of two-input AND ribocomputing devices in different strains a , An early two-input AND gate was constructed from a standard toehold switch by dividing the trigger evenly into two 15-nt domains A1 and A2. Overlap domains u and u* were designed to cause the two input RNAs to hybridize and form an active trigger. b , A domain u’ was used to vary the region complementary to u* and measure its effect on expression levels. ON/OFF GFP ratios (left axis) vary as a function of the u’ domain length. The onset of substantial GFP expression coincides with the melting temperature of u’-u* hybridization rising above 37°C (right axis). c-f , Comparison of two-input AND ribocomputing devices in RNase-deficient E. coli <t>BL21</t> Star <t>DE3</t> and non-RNase-deficient E. coli BL21 DE3. c-d , ON/OFF GFP on linear (c) and logarithmic (d) scales measured for the two-input AND gate from Figure 2e–h . e-f , ON/OFF GFP on linear (e) and logarithmic (f) scales measured for a second two-input AND gate with an identical design but different RNA sequences.
    Bl21 Star De3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 star de3/product/Thermo Fisher
    Average 99 stars, based on 57 article reviews
    Price from $9.99 to $1999.99
    bl21 star de3 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Complex cellular logic computation using ribocomputing devices"

    Article Title: Complex cellular logic computation using ribocomputing devices

    Journal: Nature

    doi: 10.1038/nature23271

    Systematic study of AND gate circuit overlap domain lengths and comparison of two-input AND ribocomputing devices in different strains a , An early two-input AND gate was constructed from a standard toehold switch by dividing the trigger evenly into two 15-nt domains A1 and A2. Overlap domains u and u* were designed to cause the two input RNAs to hybridize and form an active trigger. b , A domain u’ was used to vary the region complementary to u* and measure its effect on expression levels. ON/OFF GFP ratios (left axis) vary as a function of the u’ domain length. The onset of substantial GFP expression coincides with the melting temperature of u’-u* hybridization rising above 37°C (right axis). c-f , Comparison of two-input AND ribocomputing devices in RNase-deficient E. coli BL21 Star DE3 and non-RNase-deficient E. coli BL21 DE3. c-d , ON/OFF GFP on linear (c) and logarithmic (d) scales measured for the two-input AND gate from Figure 2e–h . e-f , ON/OFF GFP on linear (e) and logarithmic (f) scales measured for a second two-input AND gate with an identical design but different RNA sequences.
    Figure Legend Snippet: Systematic study of AND gate circuit overlap domain lengths and comparison of two-input AND ribocomputing devices in different strains a , An early two-input AND gate was constructed from a standard toehold switch by dividing the trigger evenly into two 15-nt domains A1 and A2. Overlap domains u and u* were designed to cause the two input RNAs to hybridize and form an active trigger. b , A domain u’ was used to vary the region complementary to u* and measure its effect on expression levels. ON/OFF GFP ratios (left axis) vary as a function of the u’ domain length. The onset of substantial GFP expression coincides with the melting temperature of u’-u* hybridization rising above 37°C (right axis). c-f , Comparison of two-input AND ribocomputing devices in RNase-deficient E. coli BL21 Star DE3 and non-RNase-deficient E. coli BL21 DE3. c-d , ON/OFF GFP on linear (c) and logarithmic (d) scales measured for the two-input AND gate from Figure 2e–h . e-f , ON/OFF GFP on linear (e) and logarithmic (f) scales measured for a second two-input AND gate with an identical design but different RNA sequences.

    Techniques Used: Construct, Expressing, Hybridization

    Comparison of six-input OR gate ribocomputing devices measured in RNase-deficient E. coli (BL21 Star DE3) and non-RNase-deficient E. coli (BL21 DE3, MG1655Pro) a-b , ON/OFF GFP ratios measured for the device using T7 RNA polymerase in BL21 Star DE3 and BL21 DE3 cells on linear (a) and logarithmic (b) scales. Gate and input RNAs were expressed using the T7 RNA polymerase and measured 4 hours after induction with IPTG. c-d , ON/OFF GFP ratios obtained from the OR gate using E. coli RNA polymerase in MG1655Pro cells on linear (c) and logarithmic (d) scales. Gate and input RNAs were expressed using the E. coli RNA polymerase and measured 4 hours after induction of the gate RNA with IPTG. Input and decoy RNAs were expressed using a constitutive PN25 promoter.
    Figure Legend Snippet: Comparison of six-input OR gate ribocomputing devices measured in RNase-deficient E. coli (BL21 Star DE3) and non-RNase-deficient E. coli (BL21 DE3, MG1655Pro) a-b , ON/OFF GFP ratios measured for the device using T7 RNA polymerase in BL21 Star DE3 and BL21 DE3 cells on linear (a) and logarithmic (b) scales. Gate and input RNAs were expressed using the T7 RNA polymerase and measured 4 hours after induction with IPTG. c-d , ON/OFF GFP ratios obtained from the OR gate using E. coli RNA polymerase in MG1655Pro cells on linear (c) and logarithmic (d) scales. Gate and input RNAs were expressed using the E. coli RNA polymerase and measured 4 hours after induction of the gate RNA with IPTG. Input and decoy RNAs were expressed using a constitutive PN25 promoter.

    Techniques Used:

    Related Articles

    Recombinant:

    Article Title: Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport
    Article Snippet: .. These plasmids were transformed into BL21 Star™(DE3) One Shot® Chemically Competent E. coli (Invitrogen) and expression of recombinant proteins was induced by 1 mM IPTG. .. After lysis of bacteria and purification by Ni-NTA system (QIAGEN GmbH, Hilden, Germany), the His6 -tag was removed from His6 -AS-FLAG, His6 -CS-FLAG and His6 -CS-FLAG-EEVD by AcTEV™ protease (Invitrogen).

    Article Title: An engineered stable mini-protein to plug SARS-Cov-2 Spikes
    Article Snippet: .. From the screenings, the recombinant protein was successfully over-expressed in E .coli BL21(DE3) cells, resulting in a yield of 70 mg of pure protein from one litre of bacterial culture. .. Briefly, an overnight starting culture of 10 mL was prepared for growth in 1L of Luria Bertani (LB) medium containing 50µg L−1 kanamycin, which was then induced with 0.8 mM of IPTG at 16°C for approximately 16 h. The protein was purified by sonicating bacterial cells resuspended in binding buffer (300mM NaCl, 50mM Tris-HCl, 10 mM imidazole, 2.5% (v/v) glycerol, pH 7.8) containing a protease-inhibitor cocktail (Roche Diagnostics).

    Construct:

    Article Title: DNA repair enzymes ALKBH2, ALKBH3, and AlkB oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in vitro
    Article Snippet: .. Briefly, One Shot BL21 Star (DE3)pLysS E. coli cells (Invitrogen) transformed with an AlkB variant construct were grown at 37°C until OD600 had reached ∼0.4, at which point the temperature was lowered to 30°C and protein production was induced by addition of 1 mM IPTG. .. For purification, cell pellets were resuspended in lysis buffer (10 mM Tris, pH 7.3, 300 mM NaCl, 2 mM CaCl2 , 10 mM MgCl2 , 5% (v/v) glycerol, 1 mM 2-mercaptoethanol) and lysed by sonication.

    Article Title: A Prokaryotic Membrane Sculpting BAR Domain Protein
    Article Snippet: .. Inducible BdpA expression plasmids were constructed for use in S. oneidensis MR-1, M. atlanticus CP1, and E. coli BL21(DE3) using the pBBR1-mcs2 backbone described previously( ). .. The Marionette sensor components (phlF promoter, consitutively expressed PhlF repressor, and yellow fluorescence protein (YFP)) cassette from pAJM.452( ) was cloned into the pBBR1-mcs2 backbone, and the YFP cassette was replaced with the gene encoding BdpA by Gibson assembly (primers in ).

    Positron Emission Tomography:

    Article Title: Molecular Characterization of Fluoroquinolone Resistance in Mycobacterium tuberculosis: Functional Analysis of gyrA Mutation at Position 74 ▿ Mutation at Position 74 ▿ †
    Article Snippet: .. The pET101/D-TOPO plasmids carrying the gyrA gene and the pET-15b plasmid carrying the gyrB gene were separately transformed into BL21 Star (DE3) One Shot E. coli (Invitrogen) by heat shock. .. The recombinant His-tagged gyrA and gyrB proteins were separately overexpressed and purified by the same procedure.

    Expressing:

    Article Title: ATG4B (Autophagin-1) Phosphorylation Modulates Autophagy *
    Article Snippet: .. Plasmid expressing LC3-PLA2 was transformed into E. coli BL21 (DE3, Invitrogen, C6010-03). .. For TAP-tagged ATG4B protein purification, HEK-293T cells transiently expressing human Atg4B with N-terminal affinity tags, including Streptavidin and Calmodulin binding peptide, were pelleted by centrifugation at 3000 × g and TAP-tagged ATG4B protein was purified using the manufacturer's protocol (InterPlay Mammalian TAP System by Agilent Technologies).

    Article Title: Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport
    Article Snippet: .. These plasmids were transformed into BL21 Star™(DE3) One Shot® Chemically Competent E. coli (Invitrogen) and expression of recombinant proteins was induced by 1 mM IPTG. .. After lysis of bacteria and purification by Ni-NTA system (QIAGEN GmbH, Hilden, Germany), the His6 -tag was removed from His6 -AS-FLAG, His6 -CS-FLAG and His6 -CS-FLAG-EEVD by AcTEV™ protease (Invitrogen).

    Article Title: A Prokaryotic Membrane Sculpting BAR Domain Protein
    Article Snippet: .. Inducible BdpA expression plasmids were constructed for use in S. oneidensis MR-1, M. atlanticus CP1, and E. coli BL21(DE3) using the pBBR1-mcs2 backbone described previously( ). .. The Marionette sensor components (phlF promoter, consitutively expressed PhlF repressor, and yellow fluorescence protein (YFP)) cassette from pAJM.452( ) was cloned into the pBBR1-mcs2 backbone, and the YFP cassette was replaced with the gene encoding BdpA by Gibson assembly (primers in ).

    Transformation Assay:

    Article Title: ATG4B (Autophagin-1) Phosphorylation Modulates Autophagy *
    Article Snippet: .. Plasmid expressing LC3-PLA2 was transformed into E. coli BL21 (DE3, Invitrogen, C6010-03). .. For TAP-tagged ATG4B protein purification, HEK-293T cells transiently expressing human Atg4B with N-terminal affinity tags, including Streptavidin and Calmodulin binding peptide, were pelleted by centrifugation at 3000 × g and TAP-tagged ATG4B protein was purified using the manufacturer's protocol (InterPlay Mammalian TAP System by Agilent Technologies).

    Article Title: Novel TPR-containing subunit of TOM complex functions as cytosolic receptor for Entamoeba mitosomal transport
    Article Snippet: .. These plasmids were transformed into BL21 Star™(DE3) One Shot® Chemically Competent E. coli (Invitrogen) and expression of recombinant proteins was induced by 1 mM IPTG. .. After lysis of bacteria and purification by Ni-NTA system (QIAGEN GmbH, Hilden, Germany), the His6 -tag was removed from His6 -AS-FLAG, His6 -CS-FLAG and His6 -CS-FLAG-EEVD by AcTEV™ protease (Invitrogen).

    Article Title: Molecular Characterization of Fluoroquinolone Resistance in Mycobacterium tuberculosis: Functional Analysis of gyrA Mutation at Position 74 ▿ Mutation at Position 74 ▿ †
    Article Snippet: .. The pET101/D-TOPO plasmids carrying the gyrA gene and the pET-15b plasmid carrying the gyrB gene were separately transformed into BL21 Star (DE3) One Shot E. coli (Invitrogen) by heat shock. .. The recombinant His-tagged gyrA and gyrB proteins were separately overexpressed and purified by the same procedure.

    Article Title: DNA repair enzymes ALKBH2, ALKBH3, and AlkB oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in vitro
    Article Snippet: .. Briefly, One Shot BL21 Star (DE3)pLysS E. coli cells (Invitrogen) transformed with an AlkB variant construct were grown at 37°C until OD600 had reached ∼0.4, at which point the temperature was lowered to 30°C and protein production was induced by addition of 1 mM IPTG. .. For purification, cell pellets were resuspended in lysis buffer (10 mM Tris, pH 7.3, 300 mM NaCl, 2 mM CaCl2 , 10 mM MgCl2 , 5% (v/v) glycerol, 1 mM 2-mercaptoethanol) and lysed by sonication.

    Variant Assay:

    Article Title: DNA repair enzymes ALKBH2, ALKBH3, and AlkB oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine and 5-carboxylcytosine in vitro
    Article Snippet: .. Briefly, One Shot BL21 Star (DE3)pLysS E. coli cells (Invitrogen) transformed with an AlkB variant construct were grown at 37°C until OD600 had reached ∼0.4, at which point the temperature was lowered to 30°C and protein production was induced by addition of 1 mM IPTG. .. For purification, cell pellets were resuspended in lysis buffer (10 mM Tris, pH 7.3, 300 mM NaCl, 2 mM CaCl2 , 10 mM MgCl2 , 5% (v/v) glycerol, 1 mM 2-mercaptoethanol) and lysed by sonication.

    other:

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

    Plasmid Preparation:

    Article Title: ATG4B (Autophagin-1) Phosphorylation Modulates Autophagy *
    Article Snippet: .. Plasmid expressing LC3-PLA2 was transformed into E. coli BL21 (DE3, Invitrogen, C6010-03). .. For TAP-tagged ATG4B protein purification, HEK-293T cells transiently expressing human Atg4B with N-terminal affinity tags, including Streptavidin and Calmodulin binding peptide, were pelleted by centrifugation at 3000 × g and TAP-tagged ATG4B protein was purified using the manufacturer's protocol (InterPlay Mammalian TAP System by Agilent Technologies).

    Article Title: Molecular Characterization of Fluoroquinolone Resistance in Mycobacterium tuberculosis: Functional Analysis of gyrA Mutation at Position 74 ▿ Mutation at Position 74 ▿ †
    Article Snippet: .. The pET101/D-TOPO plasmids carrying the gyrA gene and the pET-15b plasmid carrying the gyrB gene were separately transformed into BL21 Star (DE3) One Shot E. coli (Invitrogen) by heat shock. .. The recombinant His-tagged gyrA and gyrB proteins were separately overexpressed and purified by the same procedure.

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    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
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    bl21 cells - by Bioz Stars, 2020-09
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    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
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    e coli bl21 cells - by Bioz Stars, 2020-09
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    Thermo Fisher e coli strain bl21 star de3
    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 Strain Bl21 Star De3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli strain bl21 star de3/product/Thermo Fisher
    Average 89 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    e coli strain bl21 star de3 - by Bioz Stars, 2020-09
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    Image Search Results


    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