bl21 star cells  (Thermo Fisher)


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    Structured Review

    Thermo Fisher bl21 star cells
    Bl21 Star Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bl21 star cells/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bl21 star cells - by Bioz Stars, 2020-02
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: The pET28a-PUF60-His was prepared by subcloning a Bam HI-Eco RI fragment of the pcDNA3.1-PUF60 vector (cloning primers 5′-caa gat ggc gac ggc gac c and 5′-gag agg gac cac tgt cac g). .. Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: .. These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. Either a half- or full-liter of media was inoculated and grown at 37°C for 8–12 h. Expression at 18°C continued for 20–24 h, and then the cells were harvested, often frozen overnight, and resuspended in 20 mM Tris, pH 7.5, 30 mM Imidazole and 1.0 M NaCl.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: The PCR product was cloned into the pET101 vector (Invitrogen). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: Briefly, GlmU was cloned into a PET 21b vector (Novagen) to form a C -terminal (His)6 -tagged fusion protein. .. The expression was carried out in BL21 star cells (Invitrogen).

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: Paragraph title: Cloning and Recombinant Expression of PpAOC1 and PpAOC2 ... For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen).

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: .. T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen). .. Since these fusion proteins contain six-His tags, Ni-nitrilotriacetic acid agarose (QIAGEN) was used for their purification.

    Centrifugation:

    Article Title: Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni
    Article Snippet: The protein was expressed by inoculating LB with overnight transformations of BL21 star cells (ThermoFisher) with Cj0843-pET28b. .. Four hours after induction, the cultures were harvested by centrifugation and resuspended in lysis buffer containing 50mM sodium phosphate pH 8.0, 300 mM sodium chloride, and 20mM imidazole.

    Amplification:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: The genes for each expressed I-AniI variant were either amplified from clones saved from the selection process or assembled de novo ( ). .. These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ).

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Recombinant Proteins Murine PrP cDNA was amplified by PCR with a 5′-His6 tag and tobacco etch virus cleavage sequence (MRGSHHHHHHGENLYFQG) and with a 3′-Myc tag and stop codon (EQKLISEEDL). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Activity of the expressed endonucleases was measured with in vitro cleavage assays ( , ), using the target-site arrays amplified from the pCcdB plasmids as substrates.

    Construct:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Expression and purification of recombinant proteins The pET28a-PTB-His construct was generously provided by D. Black (UCLA). .. Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Constructs were generated for murine PrP23–230 , N1 (residues 23–111), and C1 (residues 112–230). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: .. T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen). .. Since these fusion proteins contain six-His tags, Ni-nitrilotriacetic acid agarose (QIAGEN) was used for their purification.

    Variant Assay:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: The genes for each expressed I-AniI variant were either amplified from clones saved from the selection process or assembled de novo ( ). .. These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ).

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: .. A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen). .. Expression of the recombinant protein was induced by adding isopropyl thiogalactoside (IPTG) to a final concentration of 1 mmol l–1 when the bacterial cultures reached an optical density at 600 nm (OD600 ) between 0.4 and 0.6.

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Incubation:

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4. .. The cleared lysates were incubated for 1 h with 5 mL of pre-equilibrated amylose resin (New England Biolabs) or glutathione agarose beads (Macherey-Nagel).

    Activity Assay:

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Paragraph title: Characterizing activity of putative endonucleases ... Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography.

    Expressing:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: .. Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight. .. Pt UGEc was purified from the supernatant of lysed cell pellets using HIS-Select Nickel Affinity Gel purification (Sigma-Aldrich).

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Paragraph title: Expression and purification of recombinant proteins ... Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. Either a half- or full-liter of media was inoculated and grown at 37°C for 8–12 h. Expression at 18°C continued for 20–24 h, and then the cells were harvested, often frozen overnight, and resuspended in 20 mM Tris, pH 7.5, 30 mM Imidazole and 1.0 M NaCl.

    Article Title: Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni
    Article Snippet: Paragraph title: Protein expression and purification ... The protein was expressed by inoculating LB with overnight transformations of BL21 star cells (ThermoFisher) with Cj0843-pET28b.

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Paragraph title: Plasmids, recombinant protein expression, and purification ... Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached.

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: Paragraph title: Protein expression, purification, and pull-down assays ... BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4.

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: .. A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen). .. Expression of the recombinant protein was induced by adding isopropyl thiogalactoside (IPTG) to a final concentration of 1 mmol l–1 when the bacterial cultures reached an optical density at 600 nm (OD600 ) between 0.4 and 0.6.

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: .. The expression was carried out in BL21 star cells (Invitrogen). .. The protein was purified by a Ni-Sepharose 6 Fast Flow column (GE Health) following a standard procedure.

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: .. For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen). ..

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: .. C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer. ..

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: To facilitate expression, maltose-binding protein (MBP) with an N-terminal His-tag was fused upstream of each endonuclease. .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography.

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: Paragraph title: Expression and purification of recombinant TAT-Foxf1 DN and TAT-Engrail proteins. ... T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen).

    Western Blot:

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen). .. Elution fractions were analyzed using GelCode blue stain reagent (Pierce) or by Western blotting using T7 antibody.

    Transformation Assay:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: .. Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C. .. Protein expression was induced with 0.25 mM IPTG for 3 h. Pellets were resuspended in a lysis buffer (50 mM Tris, pH 8.0, 300 mM NaCl, 1 mM DTT) and sonicated.

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: .. For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen). ..

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Gel Purification:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight. .. Pt UGEc was purified from the supernatant of lysed cell pellets using HIS-Select Nickel Affinity Gel purification (Sigma-Aldrich).

    Flow Cytometry:

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: The expression was carried out in BL21 star cells (Invitrogen). .. The protein was purified by a Ni-Sepharose 6 Fast Flow column (GE Health) following a standard procedure.

    Protease Inhibitor:

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 400 mM NaCl, 1 mM DTT, 10% glycerol) supplemented with EDTA-free protease inhibitor (Merck), 1 mg/mL lysozyme, and 5 μg/mL DNase I and lysed by sonication.

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 200 mM NaCl, 1 mM DTT) supplemented with EDTA-free protease inhibitor (Roche), 1 mg/mL lysozyme, and 5 μg/mL DNase I and then lysed by sonication.

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer. .. Growth was allowed to continue at 30°C for a further 3 h before the cells were pelleted and lysed by sonication in pH 7.0 urea buffer (8 M urea, 0.01 M Tris, 0.1 M NaH2 PO4 , and 25 mM complete protease inhibitor).

    Generated:

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Constructs were generated for murine PrP23–230 , N1 (residues 23–111), and C1 (residues 112–230). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Sequencing:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: .. These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. Either a half- or full-liter of media was inoculated and grown at 37°C for 8–12 h. Expression at 18°C continued for 20–24 h, and then the cells were harvested, often frozen overnight, and resuspended in 20 mM Tris, pH 7.5, 30 mM Imidazole and 1.0 M NaCl.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Recombinant Proteins Murine PrP cDNA was amplified by PCR with a 5′-His6 tag and tobacco etch virus cleavage sequence (MRGSHHHHHHGENLYFQG) and with a 3′-Myc tag and stop codon (EQKLISEEDL). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: The fusion sequence and all additional sequence modifications are detailed in the supplemental information. .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography.

    Sonication:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C. .. Protein expression was induced with 0.25 mM IPTG for 3 h. Pellets were resuspended in a lysis buffer (50 mM Tris, pH 8.0, 300 mM NaCl, 1 mM DTT) and sonicated.

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. The cells were resuspended with lysozyme and sonicated, and proteins were isolated from the soluble fraction with nickel affinity chromatography and elution with 20 mM Tris, pH 7.5, 500 mM Imidazole and 500 mM NaCl.

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 400 mM NaCl, 1 mM DTT, 10% glycerol) supplemented with EDTA-free protease inhibitor (Merck), 1 mg/mL lysozyme, and 5 μg/mL DNase I and lysed by sonication.

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 200 mM NaCl, 1 mM DTT) supplemented with EDTA-free protease inhibitor (Roche), 1 mg/mL lysozyme, and 5 μg/mL DNase I and then lysed by sonication.

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen). .. To lyse the bacterial cells and solubilize the recombinant protein, pellets were resuspended in 20 mmol l–1 phosphate buffer, 0.5 mol l–1 NaCl, 6 mol l–1 guanidine, pH 7.8, followed by a 1-min sonication.

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer. .. Growth was allowed to continue at 30°C for a further 3 h before the cells were pelleted and lysed by sonication in pH 7.0 urea buffer (8 M urea, 0.01 M Tris, 0.1 M NaH2 PO4 , and 25 mM complete protease inhibitor).

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Recombinant:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Paragraph title: Expression and purification of recombinant proteins ... Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Paragraph title: Recombinant Proteins ... Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Paragraph title: Plasmids, recombinant protein expression, and purification ... Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached.

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: Paragraph title: Recombinant protein expression and purification ... A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen).

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: Paragraph title: Cloning and Recombinant Expression of PpAOC1 and PpAOC2 ... For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen).

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: Paragraph title: Expression and purification of recombinant TAT-Foxf1 DN and TAT-Engrail proteins. ... T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen).

    Nucleic Acid Electrophoresis:

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Proteins were stored in 20 mM Tris, pH 7.5, 500 mM NaCl, 50% (v/v) glycerol; purity was assessed with sodiumdodecyl sulphate-polyacrylamide gel electrophoresis, and the concentration was determined by absorbance at 280 nm collected with a NanoDrop.

    Mutagenesis:

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: These clones were used as templates for site-directed mutagenesis PCR (AOC1_F140V and AOC1_F29I/F140V) using the Pfu polymerase. .. For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen).

    Isolation:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. The cells were resuspended with lysozyme and sonicated, and proteins were isolated from the soluble fraction with nickel affinity chromatography and elution with 20 mM Tris, pH 7.5, 500 mM Imidazole and 500 mM NaCl.

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. Proteins were isolated from the soluble fraction with nickel affinity chromatography.

    Subcloning:

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: The pET28a-PUF60-His was prepared by subcloning a Bam HI-Eco RI fragment of the pcDNA3.1-PUF60 vector (cloning primers 5′-caa gat ggc gac ggc gac c and 5′-gag agg gac cac tgt cac g). .. Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Purification:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: Paragraph title: Expression and purification of His-Pt UGEc ... Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight.

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Paragraph title: Expression and purification of recombinant proteins ... Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: Proteins were expressed in BL21 Star cells (Invitrogen). .. PrP23–230 and C1 were recovered from inclusion bodies and purified as described previously ( ).

    Article Title: Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni
    Article Snippet: Paragraph title: Protein expression and purification ... The protein was expressed by inoculating LB with overnight transformations of BL21 star cells (ThermoFisher) with Cj0843-pET28b.

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Paragraph title: Plasmids, recombinant protein expression, and purification ... Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached.

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: Paragraph title: Protein expression, purification, and pull-down assays ... BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4.

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: Paragraph title: Recombinant protein expression and purification ... A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen).

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: The procedures for the expression and purification of GlmU were described previously. .. The expression was carried out in BL21 star cells (Invitrogen).

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: .. Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Proteins were stored in 20 mM Tris, pH 7.5, 500 mM NaCl, 50% (v/v) glycerol; purity was assessed with sodiumdodecyl sulphate-polyacrylamide gel electrophoresis, and the concentration was determined by absorbance at 280 nm collected with a NanoDrop.

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: Paragraph title: Expression and purification of recombinant TAT-Foxf1 DN and TAT-Engrail proteins. ... T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen).

    Polymerase Chain Reaction:

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: The PCR product was cloned into the pET101 vector (Invitrogen). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: These clones were used as templates for site-directed mutagenesis PCR (AOC1_F140V and AOC1_F29I/F140V) using the Pfu polymerase. .. For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen).

    Affinity Chromatography:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ). .. The cells were resuspended with lysozyme and sonicated, and proteins were isolated from the soluble fraction with nickel affinity chromatography and elution with 20 mM Tris, pH 7.5, 500 mM Imidazole and 500 mM NaCl.

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. Proteins were isolated from the soluble fraction with nickel affinity chromatography.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Proteins were stored in 20 mM Tris, pH 7.5, 500 mM NaCl, 50% (v/v) glycerol; purity was assessed with sodiumdodecyl sulphate-polyacrylamide gel electrophoresis, and the concentration was determined by absorbance at 280 nm collected with a NanoDrop.

    Lysis:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight. .. Lysis of bacterial cells took place using CelLytic B 2X containing 0.2 mg/ml lysozyme, 50 U/ml benzonase (all Sigma-Aldrich) and proteinase inhibitor cocktail (Roche).

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C. .. Protein expression was induced with 0.25 mM IPTG for 3 h. Pellets were resuspended in a lysis buffer (50 mM Tris, pH 8.0, 300 mM NaCl, 1 mM DTT) and sonicated.

    Article Title: Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni
    Article Snippet: The protein was expressed by inoculating LB with overnight transformations of BL21 star cells (ThermoFisher) with Cj0843-pET28b. .. Four hours after induction, the cultures were harvested by centrifugation and resuspended in lysis buffer containing 50mM sodium phosphate pH 8.0, 300 mM sodium chloride, and 20mM imidazole.

    Article Title: An unusual arrangement of two 14-3-3-like domains in the SMG5\u2013SMG7 heterodimer is required for efficient nonsense-mediated mRNA decay
    Article Snippet: Ce SMG5 and SMG7 were coexpressed in BL21 star cells (Invitrogen) harboring plasmids encoding GST- Ce SMG5 (1–420) and His6 - Ce SMG7 1–395 grown at 37°C in ZY autoinduction medium ( ) until OD600 = 0.3 was reached. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 400 mM NaCl, 1 mM DTT, 10% glycerol) supplemented with EDTA-free protease inhibitor (Merck), 1 mg/mL lysozyme, and 5 μg/mL DNase I and lysed by sonication.

    Article Title: The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase complex to mRNAs containing nonsense codons via interaction with POP2
    Article Snippet: BL21 star cells (Invitrogen) harboring plasmids encoding GST, GST-POP2, or MBP-SMG7 732–1091 were grown at 37°C in LB medium until reaching OD600 = 0.4. .. After harvest, cells were resuspended in lysis buffer (50 mM HEPES at pH 7.5, 200 mM NaCl, 1 mM DTT) supplemented with EDTA-free protease inhibitor (Roche), 1 mg/mL lysozyme, and 5 μg/mL DNase I and then lysed by sonication.

    Article Title: Mining Endonuclease Cleavage Determinants in Genomic Sequence Data *
    Article Snippet: Expression and Purification of Proteins Genes for each homologue-based variant of I-AniI were assembled, cloned, sequence verified, and transformed into BL21 Star cells (Invitrogen). .. A half-liter or 1 liter culture of autoinduction media ( ) was then inoculated and grown at 37 °C for 8–12 h until approximate saturation, after which expression at 18 °C was allowed for 20–24 h. Cells were then harvested and resuspended in 20 mm Tris, pH 7.5, 30 mm imidazole, and 1.0 m NaCl prior to lysis via a freeze-thaw cycle, sonication, and the addition of lysozyme.

    Plasmid Preparation:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: Expression and purification of His-Pt UGEc Pt UGEc was introduced into pDEST17 expression vector (Invitrogen) containing an N-terminal 6xHis-tag and an IPTG inducible promoter. .. Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight.

    Article Title: Allele-specific recognition of the 3? splice site of INS intron 1
    Article Snippet: The pET28a-PUF60-His was prepared by subcloning a Bam HI-Eco RI fragment of the pcDNA3.1-PUF60 vector (cloning primers 5′-caa gat ggc gac ggc gac c and 5′-gag agg gac cac tgt cac g). .. Following transformation, BL21 Star cells (Invitrogen) were grown to optical density of 0.8 at 37°C.

    Article Title: An N-terminal Fragment of the Prion Protein Binds to Amyloid-β Oligomers and Inhibits Their Neurotoxicity in Vivo *
    Article Snippet: The PCR product was cloned into the pET101 vector (Invitrogen). .. Proteins were expressed in BL21 Star cells (Invitrogen).

    Article Title: Structural studies and molecular dynamics simulations suggest a processive mechanism of exolytic lytic transglycosylase from Campylobacter jejuni
    Article Snippet: Protein expression and purification The Cj0843 expression vector (Cj0843-pET28b) was used for expression as previously described [ ]. .. The protein was expressed by inoculating LB with overnight transformations of BL21 star cells (ThermoFisher) with Cj0843-pET28b.

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: .. A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen). .. Expression of the recombinant protein was induced by adding isopropyl thiogalactoside (IPTG) to a final concentration of 1 mmol l–1 when the bacterial cultures reached an optical density at 600 nm (OD600 ) between 0.4 and 0.6.

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: Briefly, GlmU was cloned into a PET 21b vector (Novagen) to form a C -terminal (His)6 -tagged fusion protein. .. The expression was carried out in BL21 star cells (Invitrogen).

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: .. For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen). ..

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: .. C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer. ..

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Endonuclease genes were transferred from the pENDO-HE plasmid into the pET15-HE plasmid for protein expression. .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography.

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: .. T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen). .. Since these fusion proteins contain six-His tags, Ni-nitrilotriacetic acid agarose (QIAGEN) was used for their purification.

    Selection:

    Article Title: Reprogramming homing endonuclease specificity through computational design and directed evolution
    Article Snippet: The genes for each expressed I-AniI variant were either amplified from clones saved from the selection process or assembled de novo ( ). .. These genes were cloned into pET15-HE with NcoI and NotI site, sequence verified and expressed in BL21 Star cells (Invitrogen) using autoinduction ( ).

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: A single round of selection was completed, followed by collection of the plasmids and retransformation to obtain more accurate values for bacteria survival. .. Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography.

    In Vitro:

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: Paragraph title: His-tagged protein preparation and in vitro kinase assays. ... C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Activity of the expressed endonucleases was measured with in vitro cleavage assays ( , ), using the target-site arrays amplified from the pCcdB plasmids as substrates.

    Concentration Assay:

    Article Title: A gene stacking approach leads to engineered plants with highly increased galactan levels in Arabidopsis
    Article Snippet: .. Gene expression in BL21 Star cells (Invitrogen) was induced by adding IPTG to a final concentration of 1 mM, and cultures were grown at 18°C overnight. .. Pt UGEc was purified from the supernatant of lysed cell pellets using HIS-Select Nickel Affinity Gel purification (Sigma-Aldrich).

    Article Title: Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis
    Article Snippet: A partial cDNA encoding the C-terminal 155 amino acids of a variant of the TMP family was subcloned into the pRSET expression vector (Invitrogen) and subsequently expressed in BL21 Star cells (Invitrogen). .. Expression of the recombinant protein was induced by adding isopropyl thiogalactoside (IPTG) to a final concentration of 1 mmol l–1 when the bacterial cultures reached an optical density at 600 nm (OD600 ) between 0.4 and 0.6.

    Article Title: Crystal Structures of Physcomitrella patens AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] AOC1 and AOC2: Insights into the Enzyme Mechanism and Differences in Substrate Specificity 1 [W] [OA]
    Article Snippet: For expression, the respective plasmid was transformed in BL21 Star cells (Invitrogen). .. Protein expression was induced with a final concentration of 1 m m isopropyl-β- d -thiogalactopyranoside.

    Article Title: Massively parallel determination and modeling of endonuclease substrate specificity
    Article Snippet: Proteins were expressed in BL21 Star cells (Invitrogen) using a half-liter of media and autoinduction ( ) and purified with nickel affinity chromatography. .. Proteins were stored in 20 mM Tris, pH 7.5, 500 mM NaCl, 50% (v/v) glycerol; purity was assessed with sodiumdodecyl sulphate-polyacrylamide gel electrophoresis, and the concentration was determined by absorbance at 280 nm collected with a NanoDrop.

    Staining:

    Article Title: Forkhead Box F1 Is Essential for Migration of Mesenchymal Cells and Directly Induces Integrin-Beta3 Expression ▿
    Article Snippet: T7-Foxf1 DN or T7-Engrail constructs were cloned into the pTAT vector, which was used to transform BL21 Star cells (Invitrogen). .. Elution fractions were analyzed using GelCode blue stain reagent (Pierce) or by Western blotting using T7 antibody.

    Positron Emission Tomography:

    Article Title: Chemoenzymatic synthesis of Uridine Diphosphate-GlcNAc and Uridine Diphosphate-GalNAc analogs for the preparation of unnatural glycosaminoglycans
    Article Snippet: Briefly, GlmU was cloned into a PET 21b vector (Novagen) to form a C -terminal (His)6 -tagged fusion protein. .. The expression was carried out in BL21 star cells (Invitrogen).

    Article Title: Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿Phosphorylation of the Human Papillomavirus Type 16 E1^E4 Protein at T57 by ERK Triggers a Structural Change That Enhances Keratin Binding and Protein Stability ▿ †
    Article Snippet: .. C-terminally hexahistidine-tagged 16E1^E4 with a leucine-glutamic acid linker (His-16E1^E4) was expressed from the pET-28(b)+/16E1^E4 expression vector in BL21 Star cells (Invitrogen, United Kingdom) as described by the manufacturer. ..

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