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Stratagene e coli xl1 blue strain
E Coli Xl1 Blue Strain, supplied by Stratagene, used in various techniques. Bioz Stars score: 96/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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e coli xl1 blue strain - by Bioz Stars, 2020-04
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Transduction:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Transformation of final S. aureus strains with pMAD and pSD3-3 plasmids were performed by phage transduction as described previously .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Transformation of final S. aureus strains with pMAD and pSD3-3 plasmids were performed by phage transduction as described previously .

Clone Assay:

Article Title: Replication Stalling at Friedreich's Ataxia (GAA)n Repeats In Vivo
Article Snippet: .. Cloning was carried out in the Escherichia coli XL1-Blue strain (Stratagene). .. Yeast replication studies were performed in S. cerevisiae CH1585 ( MAT a leu2 Δ 1 trp Δ 63 ura3-52 his3-200 ) strain (ATCC 96098).

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene). ..

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene). .. Protein Expression and Purification FP-His12 proteins were expressed in E. coli BL21 Star (DE3) strain (Invitrogen).

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies. .. It was grown in standard Luria-Bertani medium (Pronadisa) at 37°C using a final concentration of 100 μg ml−1 of ampicillin for the selection of the pGEM-T Easy plasmid (Promega).

Centrifugation:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Following lysis by sonication (1 pulse) for 5 s on ice, cytosol was obtained by centrifugation at 100,000 g for 30 min at 4°C, and analyzed with cGMP and cAMP (Sigma). .. YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Cells harvested by centrifugation (1,500 g , 15 min) were washed with 20 ml of 10 mM Tris-HCl (pH 8.0) containing 0.1 M NaCl and 1 mM EDTA, and suspended with 10 ml of 25 mM Tris-HCl (pH 8.0) containing 50 mM glucose and 10 mM EDTA.

Amplification:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol. .. Recombinant fusion peptide of the human COL17 NC16A domain (rhNC16A) with glutathione S-transferase (GST) was synthesized as reported previously.

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: Genes for mCherry and mGFP were amplified by PCR and cloned into the Nco I/Hin dIII sites of pET-28b(+)-His12. mCherry-mGFP-His12 was constructed sequentially by first cloning mCherry into the Nco I/Bam HI sites of pET28b(+) with an oligo cassette encoding a 12× His-tag downstream of the fluorescent protein to generate pET-28b(+)-mCherry1 -His12. mGFP was then inserted into the Bam HI/Hin dIII sites of pET-28b(+)-mCherry1 -His12 to produce pET-28b(+)-mCherry1 -mGFP2 -His12. .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene).

DNA Ligation:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: FastDigest restriction enzymes and Rapid DNA ligation kit (Thermo Scientific) were used according to the manufacturer’s instructions. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: FastDigest restriction enzymes and Rapid DNA ligation kit (Thermo Scientific) were used according to the manufacturer’s instructions. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: FastDigest restriction enzymes and Rapid DNA ligation kit (Thermo Scientific) were used according to the manufacturer’s instructions. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: FastDigest restriction enzymes and Rapid DNA ligation kit (Thermo Scientific) were used according to the manufacturer’s instructions. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Construct:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: The phage antibody libraries were constructed by randomly combining the genes coding Fd fragments of IgG heavy chains with IgG light chain genes of either lambda or kappa DNA in equal amounts (see Supplemental Figure S1 at http://ajp.amjpathol.org.). .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: This construct was called EC1–2TM and was cloned into the KpnI and SpeI sites of the aforementioned expression vector, which contains the I27 marker domains ( ). .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene).

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: Genes for mCherry and mGFP were amplified by PCR and cloned into the Nco I/Hin dIII sites of pET-28b(+)-His12. mCherry-mGFP-His12 was constructed sequentially by first cloning mCherry into the Nco I/Bam HI sites of pET28b(+) with an oligo cassette encoding a 12× His-tag downstream of the fluorescent protein to generate pET-28b(+)-mCherry1 -His12. mGFP was then inserted into the Bam HI/Hin dIII sites of pET-28b(+)-mCherry1 -His12 to produce pET-28b(+)-mCherry1 -mGFP2 -His12. .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene).

Enzyme-linked Immunosorbent Assay:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: The diagnosis of BP was made by the typical clinical and histological manifestations as well as by laboratory data including anti-COL17 ELISA and indirect immunofluorescence (IIF). .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Incubation:

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: .. Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Cells harvested by centrifugation (1,500 g , 15 min) were washed with 20 ml of 10 mM Tris-HCl (pH 8.0) containing 0.1 M NaCl and 1 mM EDTA, and suspended with 10 ml of 25 mM Tris-HCl (pH 8.0) containing 50 mM glucose and 10 mM EDTA.

Expressing:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Paragraph title: Protein expression, in vitro spectroscopy and pH titrations ... YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Phagemid expression vector p3MH, a gift from Dr. Yan Wang (Central Lab of Navy General Hospital, Beijing, China), was derived from pCOMB3H (Scripps Research Institute, La Jolla, CA) by adding 9E10/c- myc epitope for detection and a hexahistidine tag for column purification at the 3′ end of Fd. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: .. E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The E. coli BL21(DE3) strain (F− ompT gal [dcm] [lon] hsdSB (rB − mB − ) (λ DE3)) was employed in MIC determination and protein purification.

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: For expression in eukaryotic cells, a PCR-amplified Age I– Bgl II fragment encoding TurboGFP or ppluGFP2 was swapped with EGFP in pEGFP-C1 vector (Clontech), resulting in TurboGFP-C1 or ppluGFP2-C1 plasmid. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Article Title: Treatment with p33 Curtails Morbidity and Mortality in a Histone-Induced Murine Shock Model
Article Snippet: .. The pMAL-c2 expression vector (New England Biolabs, Ipswich, Mass., USA) and E. coli XL1-Blue strain (Stratagene, Heidelberg, Germany) were used to express recombinant MBP-p33. .. Purification and removal of the MBP-tag was performed as previously described [ ].

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: This construct was called EC1–2TM and was cloned into the KpnI and SpeI sites of the aforementioned expression vector, which contains the I27 marker domains ( ). .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene).

Transformation Assay:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Article Title: Histidine kinases mediate differentiation, stress response, and pathogenicity in Magnaporthe oryzae
Article Snippet: Escherichia coli XL1-BLUE strain (Stratagene Products Division, La Jolla, CA) was used for routine bacterial transformations and construction of plasmids. .. Transformations of M. oryzae were conducted using Agrobacterium tumefaciens -mediated transformation (ATMT; De Groot et al. ; Rho et al. ).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Derivative Assay:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Phagemid expression vector p3MH, a gift from Dr. Yan Wang (Central Lab of Navy General Hospital, Beijing, China), was derived from pCOMB3H (Scripps Research Institute, La Jolla, CA) by adding 9E10/c- myc epitope for detection and a hexahistidine tag for column purification at the 3′ end of Fd. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Electroporation:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Transfection:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Protein expression, in vitro spectroscopy and pH titrations Cygnus was transiently transfected into HEK293T cells using FuGENE 6 (Roche), and one or two days after transfection, cells were washed three times with chilled PBS, scraped from the plate, and resuspended in 5 mM Tris-HCl, 2 mM EDTA (pH 7.3). .. YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: Fluorescence images of transfected HeLa cells were obtained using the Olympus CK40 inverted microscope equipped with the Olympus DP50 camera. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: .. Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Cells harvested by centrifugation (1,500 g , 15 min) were washed with 20 ml of 10 mM Tris-HCl (pH 8.0) containing 0.1 M NaCl and 1 mM EDTA, and suspended with 10 ml of 25 mM Tris-HCl (pH 8.0) containing 50 mM glucose and 10 mM EDTA.

Sequencing:

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene). ..

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies. .. It was grown in standard Luria-Bertani medium (Pronadisa) at 37°C using a final concentration of 100 μg ml−1 of ampicillin for the selection of the pGEM-T Easy plasmid (Promega).

Protease Inhibitor:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene). .. Cultures were grown overnight at 37°C, and pellets were lysed by sonication in a solution of 25 mM Tris-HCl (pH 8.0), 1 mM β-mercaptoethanol and a protease inhibitor cocktail for use with bacterial cells (Sigma).

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Egg white lysozyme was added to the cell suspension at a final concentration of 100 μg/ml, and the mixture was incubated for 10 min. After addition of 10 ml HSEI buffer (10 mM Hepes-NaOH [pH 7.5] containing 0.25 M sucrose, 1 mM EDTA, and a protease inhibitor cocktail [EDTA-free Complete™ Protease Inhibitor; Roche]), the resulting mixture was sonicated five times with a probe-type sonicator at 20 W for 10 s The sonicated sample was centrifuged (1500 g , 15 min), and the supernatant fluid was recovered as cell lysate.

Generated:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol . .. Staphyloccocal electrocompetent cells were generated as previously described .

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Article Title: Histidine kinases mediate differentiation, stress response, and pathogenicity in Magnaporthe oryzae
Article Snippet: Escherichia coli XL1-BLUE strain (Stratagene Products Division, La Jolla, CA) was used for routine bacterial transformations and construction of plasmids. .. Magnaporthe oryzae -mutant strains were generated using a hygromycin resistance (hygromycin-phospho-transferase gene, HPT ; Odenbach et al. ) or a glufosinate-ammonium resistance (phosphinothricin-acetyl-transferase gene, BAR ; Kramer et al. ).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ). .. Staphylococcal electrocompetent cells were generated as previously described ( ).

Imaging:

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: Expression in mammalian cell lines and imaging. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Inverted Microscopy:

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: Fluorescence images of transfected HeLa cells were obtained using the Olympus CK40 inverted microscope equipped with the Olympus DP50 camera. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Using previously described methods and a set of PCR primers , , , antibody genes were amplified by RT-PCR from approximately 1 × 108 mononuclear cells isolated from 50 ml of peripheral blood from each patient. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Sonication:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Following lysis by sonication (1 pulse) for 5 s on ice, cytosol was obtained by centrifugation at 100,000 g for 30 min at 4°C, and analyzed with cGMP and cAMP (Sigma). .. YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Egg white lysozyme was added to the cell suspension at a final concentration of 100 μg/ml, and the mixture was incubated for 10 min. After addition of 10 ml HSEI buffer (10 mM Hepes-NaOH [pH 7.5] containing 0.25 M sucrose, 1 mM EDTA, and a protease inhibitor cocktail [EDTA-free Complete™ Protease Inhibitor; Roche]), the resulting mixture was sonicated five times with a probe-type sonicator at 20 W for 10 s The sonicated sample was centrifuged (1500 g , 15 min), and the supernatant fluid was recovered as cell lysate.

Recombinant:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol. .. Recombinant fusion peptide of the human COL17 NC16A domain (rhNC16A) with glutathione S-transferase (GST) was synthesized as reported previously.

Article Title: Treatment with p33 Curtails Morbidity and Mortality in a Histone-Induced Murine Shock Model
Article Snippet: .. The pMAL-c2 expression vector (New England Biolabs, Ipswich, Mass., USA) and E. coli XL1-Blue strain (Stratagene, Heidelberg, Germany) were used to express recombinant MBP-p33. .. Purification and removal of the MBP-tag was performed as previously described [ ].

Immunofluorescence:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: The diagnosis of BP was made by the typical clinical and histological manifestations as well as by laboratory data including anti-COL17 ELISA and indirect immunofluorescence (IIF). .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Transmission Electron Microscopy:

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The ompA-TEM-1 fusion in pET-TEM-1 also contains an E28G substitution in TEM-1 near the signal cleavage site.

Fluorescence:

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: Fluorescence images of transfected HeLa cells were obtained using the Olympus CK40 inverted microscope equipped with the Olympus DP50 camera. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Mutagenesis:

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: .. E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The E. coli BL21(DE3) strain (F− ompT gal [dcm] [lon] hsdSB (rB − mB − ) (λ DE3)) was employed in MIC determination and protein purification.

Article Title: Histidine kinases mediate differentiation, stress response, and pathogenicity in Magnaporthe oryzae
Article Snippet: Escherichia coli XL1-BLUE strain (Stratagene Products Division, La Jolla, CA) was used for routine bacterial transformations and construction of plasmids. .. Magnaporthe oryzae -mutant strains were generated using a hygromycin resistance (hygromycin-phospho-transferase gene, HPT ; Odenbach et al. ) or a glufosinate-ammonium resistance (phosphinothricin-acetyl-transferase gene, BAR ; Kramer et al. ).

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: To construct the triple mutant (TM) D67A/D100A/D134A, the wild type EC1–2 sequence was cloned into the pT7blue vector (Novagen) using KpnI and SpeI sites prior to performing site-directed mutagenesis (one point mutation at a time) using the QuikChange kit (Stratagene). .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene).

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: L. plantarum NCIMB8826 and its l -arabinose isomerase-deficient mutant BPT197 were cultivated at 30°C in standard MRS growth medium (Lab M Limited) and in simplified MRS medium in bioreactor cultivations. .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies.

Isolation:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Using previously described methods and a set of PCR primers , , , antibody genes were amplified by RT-PCR from approximately 1 × 108 mononuclear cells isolated from 50 ml of peripheral blood from each patient. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Article Title: Histidine kinases mediate differentiation, stress response, and pathogenicity in Magnaporthe oryzae
Article Snippet: DNA manipulation/construction of gene inactivation vectors Genomic DNA was isolated from lyophilized mycelium of 4-day-old liquid cultures, grown at 26°C and 120 rpm, using DNeasy® Plant Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. .. Escherichia coli XL1-BLUE strain (Stratagene Products Division, La Jolla, CA) was used for routine bacterial transformations and construction of plasmids.

Purification:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene). .. Purified proteins were dialyzed into PBS (pH 7.4) using a PD-10 column (GE Healthcare). pH titrations of the absorption were performed by using the buffers containing 125 mM KCl, 20 mM NaCl, 0.5 mM CaCl2 , 0.5 mM MgCl2 and 25 mM of ethanolamine (pH 10.0), TAPS (pH 9.0, 8.5), HEPES (pH 8.0, 7.5, 7.0), MES (pH 6.5, 6.0, 5.5), or acetate (pH 5.0, 4.5, 4.0).

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol. .. Recombinant fusion peptide of the human COL17 NC16A domain (rhNC16A) with glutathione S-transferase (GST) was synthesized as reported previously.

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech). ..

Article Title: Treatment with p33 Curtails Morbidity and Mortality in a Histone-Induced Murine Shock Model
Article Snippet: Paragraph title: Purification of Recombinant Maltose-Binding Protein-p33 (MBP-p33) ... The pMAL-c2 expression vector (New England Biolabs, Ipswich, Mass., USA) and E. coli XL1-Blue strain (Stratagene, Heidelberg, Germany) were used to express recombinant MBP-p33.

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Protein Purification:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene). .. Protein purification was carried out using His GraviTrap (GE Healthcare).

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The E. coli BL21(DE3) strain (F− ompT gal [dcm] [lon] hsdSB (rB − mB − ) (λ DE3)) was employed in MIC determination and protein purification.

Polymerase Chain Reaction:

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Using previously described methods and a set of PCR primers , , , antibody genes were amplified by RT-PCR from approximately 1 × 108 mononuclear cells isolated from 50 ml of peripheral blood from each patient. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: For expression in eukaryotic cells, a PCR-amplified Age I– Bgl II fragment encoding TurboGFP or ppluGFP2 was swapped with EGFP in pEGFP-C1 vector (Clontech), resulting in TurboGFP-C1 or ppluGFP2-C1 plasmid. .. Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech).

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: Genes for mCherry and mGFP were amplified by PCR and cloned into the Nco I/Hin dIII sites of pET-28b(+)-His12. mCherry-mGFP-His12 was constructed sequentially by first cloning mCherry into the Nco I/Bam HI sites of pET28b(+) with an oligo cassette encoding a 12× His-tag downstream of the fluorescent protein to generate pET-28b(+)-mCherry1 -His12. mGFP was then inserted into the Bam HI/Hin dIII sites of pET-28b(+)-mCherry1 -His12 to produce pET-28b(+)-mCherry1 -mGFP2 -His12. .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene).

Positron Emission Tomography:

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The pET-TEM-1 vector encodes an ompA leader-TEM-1 fusion gene driven by the T7 promoter .

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: Genes for mCherry and mGFP were amplified by PCR and cloned into the Nco I/Hin dIII sites of pET-28b(+)-His12. mCherry-mGFP-His12 was constructed sequentially by first cloning mCherry into the Nco I/Bam HI sites of pET28b(+) with an oligo cassette encoding a 12× His-tag downstream of the fluorescent protein to generate pET-28b(+)-mCherry1 -His12. mGFP was then inserted into the Bam HI/Hin dIII sites of pET-28b(+)-mCherry1 -His12 to produce pET-28b(+)-mCherry1 -mGFP2 -His12. .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene).

Spectroscopy:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Paragraph title: Protein expression, in vitro spectroscopy and pH titrations ... YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Electroporation Bacterial Transformation:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Paragraph title: DNA manipulations and bacterial transformation ... Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Paragraph title: DNA Manipulations and Bacterial Transformation ... Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Plasmid Preparation:

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: Blockade of Autoantibody-Initiated Tissue Damage by Using Recombinant Fab Antibody Fragments against Pathogenic Autoantigen
Article Snippet: Phagemid expression vector p3MH, a gift from Dr. Yan Wang (Central Lab of Navy General Hospital, Beijing, China), was derived from pCOMB3H (Scripps Research Institute, La Jolla, CA) by adding 9E10/c- myc epitope for detection and a hexahistidine tag for column purification at the 3′ end of Fd. .. The phagemid libraries were electroporated into E. coli XL1-Blue strain (Stratagene, La Jolla, CA), and the phage display of the libraries was performed as described elsewhere., Before amplification, the resulting libraries were examined for the coexpression of heavy and light chains by enzyme digestion and for the diversity by fingerprinting of antibody genes (Fd and light chain) of 24 randomly selected single colonies., The amplified recombinant phages were purified from culture supernatants by polyethylene glycol precipitation and resuspended in PBS, pH 7.4, containing 1% bovine serum albumin (BSA) and 10% glycerol.

Article Title: Genetic and structural characterization of an L201P global suppressor substitution in TEM-1 ?-lactamase
Article Snippet: E. coli XL1-Blue strain ( recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F' proAB lacIq ZΔM15 Tn10 (Tetr )]) was obtained from Stratagene (La Jolla, CA) and utilized in site-directed mutagenesis and determination of steady-state protein expression levels. .. The pET-TEM-1 vector encodes an ompA leader-TEM-1 fusion gene driven by the T7 promoter .

Article Title: Sensory deprivation in Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) by electroporation and then introduced in S. aureus by electroporation using a previously described protocol .

Article Title: Treatment with p33 Curtails Morbidity and Mortality in a Histone-Induced Murine Shock Model
Article Snippet: .. The pMAL-c2 expression vector (New England Biolabs, Ipswich, Mass., USA) and E. coli XL1-Blue strain (Stratagene, Heidelberg, Germany) were used to express recombinant MBP-p33. .. Purification and removal of the MBP-tag was performed as previously described [ ].

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: This construct was called EC1–2TM and was cloned into the KpnI and SpeI sites of the aforementioned expression vector, which contains the I27 marker domains ( ). .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene).

Article Title: A Systematic Evaluation of the Two-Component Systems Network Reveals That ArlRS Is a Key Regulator of Catheter Colonization by Staphylococcus aureus
Article Snippet: Plasmids were purified using the NucleoSpin Plasmid miniprep kit (Macherey-Nagel) according to the manufacturer’s protocol. .. Plasmids were transformed into E. coli XL1-Blue strain (Stratagene) and S. aureus by electroporation, using previously described protocols ( ).

Article Title: Monitoring Lipid Anchor Organization in Cell Membranes by PIE-FCCS
Article Snippet: Cloning: Construction of His-Tagged Fluorescent Proteins (FP-His12) Genes were cloned into the Nco I/Xho I restriction sites in the multiple cloning region downstream of a T7 promoter in the vector pET-28b(+) (Novagen). .. All cloning was carried out in E. coli XL1-Blue strain (Stratagene).

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: Erythromycin was used at a concentration of 5 μg liter−1 for the selection of plasmid pRKDEL. .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies.

Selection:

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: Erythromycin was used at a concentration of 5 μg liter−1 for the selection of plasmid pRKDEL. .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies.

In Vitro:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Paragraph title: Protein expression, in vitro spectroscopy and pH titrations ... YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

Spectrophotometry:

Article Title: Structural basis for the fast maturation of Arthropoda green fluorescent protein
Article Snippet: Proteins fused to the N-terminal polyhistidine tag were expressed in E. coli XL1-blue strain (Stratagene, La Jolla, CA, USA) and purified using TALON metal-affinity resin (Clontech). .. Absorption spectra were recorded with a Beckman DU520 UV/VIS Spectrophotometer.

Concentration Assay:

Article Title: Plasmodium falciparum Phospholipase C Hydrolyzing Sphingomyelin and Lysocholinephospholipids Is a Possible Target for Malaria Chemotherapy
Article Snippet: Logarithmically growing cells of E. coli XL1-Blue strain (Stratagene) transfected with pGEX-PfNSM or pGEX-6P-2 were incubated in 250 ml Terrific broth containing 50 μg/ml ampicillin and 1 mM isopropyl-1-thio-β-d-galactopyranoside for 90 min at 33°C. .. Egg white lysozyme was added to the cell suspension at a final concentration of 100 μg/ml, and the mixture was incubated for 10 min. After addition of 10 ml HSEI buffer (10 mM Hepes-NaOH [pH 7.5] containing 0.25 M sucrose, 1 mM EDTA, and a protease inhibitor cocktail [EDTA-free Complete™ Protease Inhibitor; Roche]), the resulting mixture was sonicated five times with a probe-type sonicator at 20 W for 10 s The sonicated sample was centrifuged (1500 g , 15 min), and the supernatant fluid was recovered as cell lysate.

Article Title: Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose ▿
Article Snippet: Erythromycin was used at a concentration of 5 μg liter−1 for the selection of plasmid pRKDEL. .. Escherichia coli XL1-Blue strain (Stratagene) was used as the cloning host in sequencing studies.

Marker:

Article Title: Nanomechanics of the Cadherin Ectodomain
Article Snippet: This construct was called EC1–2TM and was cloned into the KpnI and SpeI sites of the aforementioned expression vector, which contains the I27 marker domains ( ). .. All the sequences were verified by sequencing both strands of the DNA, and the cloning steps were carried out in the Escherichia coli XL1-Blue strain (Stratagene).

Lysis:

Article Title: Blue Fluorescent cGMP Sensor for Multiparameter Fluorescence Imaging
Article Snippet: Following lysis by sonication (1 pulse) for 5 s on ice, cytosol was obtained by centrifugation at 100,000 g for 30 min at 4°C, and analyzed with cGMP and cAMP (Sigma). .. YFPs with an N-terminal polyhistidine tag were expressed in E. coli XL1-Blue strain (Stratagene).

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  • 87
    Stratagene e coli strain xl1 blue mr
    Growth of E. coli strains on (GlcNAc) 2 and cellobiose. E. coli strains <t>XL1-Blue</t> MR (wild type, •), Xm1.4 (transposon mutant, ▴), and Xm1.4:pES1 (transposon mutant harboring the 7.3-kb clone, ○) were tested for their ability to grow on (GlcNAc) 2 (solid lines) and cellobiose (dashed lines) as sole carbon sources. Minimal media (M9 salts) supplemented with 0.5 mM thiamine and 0.05% Casamino acids containing either 10 mM (GlcNAc) 2 or 10 mM cellobiose were inoculated using a 1:100 dilution of overnight cultures grown in LB. Growth was monitored over the indicated time course by using a Klett photoelectric colorimeter with a no. 54 green filter (550 nm). At the arrow, an aliquot of Xm1.4:pES1 cells [preinduced by growth to mid-logarithmic phase on (GlcNAc) 2 ] was harvested and washed three times with an equal volume of minimal medium and then resuspended in minimal medium containing 10 mM cellobiose.
    E Coli Strain Xl1 Blue Mr, supplied by Stratagene, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli strain xl1 blue mr/product/Stratagene
    Average 87 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    e coli strain xl1 blue mr - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    95
    Stratagene e coli xl1 blue
    Immunoblot showing OmpL1 expression at different IPTG concentrations. A culture of E. coli <t>XL1-Blue</t> containing plasmid pMMB66-OmpL1 was grown to log phase, separated into four equal volumes, and treated with various concentrations of IPTG. Cells were incubated for an additional 3 h in a shaker incubator at 37°C, pelleted in a microcentrifuge, resuspended in final sample buffer, and analyzed by SDS-PAGE and immunoblotting. A companion SDS-polyacrylamide gel loaded with the same four samples was stained with Coomassie brilliant blue to verify that all four lanes contained comparable amounts of material (data not shown). The locations of molecular size standards are shown (in kilodaltons) on the left. Even though use of the tac promoter typically results in relatively high background expression levels, expression using plasmid pMMB66-OmpL1 was not detectable without IPTG induction.
    E Coli Xl1 Blue, supplied by Stratagene, used in various techniques. Bioz Stars score: 95/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli xl1 blue/product/Stratagene
    Average 95 stars, based on 43 article reviews
    Price from $9.99 to $1999.99
    e coli xl1 blue - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    Image Search Results


    (A) Recombinant expression of GST-tagged BPS in E. coli XL 1-Blue MRF

    Journal: Infection and Immunity

    Article Title: Molecular Analysis of Group B Protective Surface Protein, a New Cell Surface Protective Antigen of Group B Streptococci

    doi:

    Figure Lengend Snippet: (A) Recombinant expression of GST-tagged BPS in E. coli XL 1-Blue MRF" and purification by glutathione-agarose affinity chromatography. The Coomassie-stained gel shows E. coli whole-cell extracts prior to induction (lane 1) and after induction (lane 2) and purified recombinant BPS fusion protein (lane 3). (B) Western blot analysis of whole-cell lysates of GBS strain Compton R (lane 1), E. coli XL 1-Blue MRF" harboring plasmid pGEX2T (lane 2), and E. coli XL 1-Blue MRF" harboring plasmid pSE4 (lane 3) using a polyclonal antiserum raised against purified recombinant GST-BPS fusion protein. (C) Western blot analysis of purified recombinant BPS protein as well as whole-cell extracts of GBS and E. coli using BPS antiserum. Lane 1, purified recombinant BPS; lane 2, Compton R; lane 3, 71-735 (serotype III/R1); lane 4, H4A-0126 (type Ia/R1, BPS); lane 5, 76-043 (type III/R4); lane 6, E. coli XL1-Blue expressing BPS; lane 7, E. coli XL1-Blue control.

    Article Snippet: The Escherichia coli strains XL1-Blue MRF and XLOLR were obtained from a commercial source (Stratagene).

    Techniques: Recombinant, Expressing, Purification, Affinity Chromatography, Staining, Western Blot, Plasmid Preparation, IA

    Growth of E. coli strains on (GlcNAc) 2 and cellobiose. E. coli strains XL1-Blue MR (wild type, •), Xm1.4 (transposon mutant, ▴), and Xm1.4:pES1 (transposon mutant harboring the 7.3-kb clone, ○) were tested for their ability to grow on (GlcNAc) 2 (solid lines) and cellobiose (dashed lines) as sole carbon sources. Minimal media (M9 salts) supplemented with 0.5 mM thiamine and 0.05% Casamino acids containing either 10 mM (GlcNAc) 2 or 10 mM cellobiose were inoculated using a 1:100 dilution of overnight cultures grown in LB. Growth was monitored over the indicated time course by using a Klett photoelectric colorimeter with a no. 54 green filter (550 nm). At the arrow, an aliquot of Xm1.4:pES1 cells [preinduced by growth to mid-logarithmic phase on (GlcNAc) 2 ] was harvested and washed three times with an equal volume of minimal medium and then resuspended in minimal medium containing 10 mM cellobiose.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Wild-type Escherichia coli grows on the chitin disaccharide, N,N?-diacetylchitobiose, by expressing the cel operon

    doi:

    Figure Lengend Snippet: Growth of E. coli strains on (GlcNAc) 2 and cellobiose. E. coli strains XL1-Blue MR (wild type, •), Xm1.4 (transposon mutant, ▴), and Xm1.4:pES1 (transposon mutant harboring the 7.3-kb clone, ○) were tested for their ability to grow on (GlcNAc) 2 (solid lines) and cellobiose (dashed lines) as sole carbon sources. Minimal media (M9 salts) supplemented with 0.5 mM thiamine and 0.05% Casamino acids containing either 10 mM (GlcNAc) 2 or 10 mM cellobiose were inoculated using a 1:100 dilution of overnight cultures grown in LB. Growth was monitored over the indicated time course by using a Klett photoelectric colorimeter with a no. 54 green filter (550 nm). At the arrow, an aliquot of Xm1.4:pES1 cells [preinduced by growth to mid-logarithmic phase on (GlcNAc) 2 ] was harvested and washed three times with an equal volume of minimal medium and then resuspended in minimal medium containing 10 mM cellobiose.

    Article Snippet: E. coli strain XL1-Blue MR was purchased from Stratagene, and E. coli strain LR-175 , Salmonella typhimurium strain LT2 , and other strains were obtained from the American Type Culture Collection.

    Techniques: Mutagenesis

    Immunoblot showing OmpL1 expression at different IPTG concentrations. A culture of E. coli XL1-Blue containing plasmid pMMB66-OmpL1 was grown to log phase, separated into four equal volumes, and treated with various concentrations of IPTG. Cells were incubated for an additional 3 h in a shaker incubator at 37°C, pelleted in a microcentrifuge, resuspended in final sample buffer, and analyzed by SDS-PAGE and immunoblotting. A companion SDS-polyacrylamide gel loaded with the same four samples was stained with Coomassie brilliant blue to verify that all four lanes contained comparable amounts of material (data not shown). The locations of molecular size standards are shown (in kilodaltons) on the left. Even though use of the tac promoter typically results in relatively high background expression levels, expression using plasmid pMMB66-OmpL1 was not detectable without IPTG induction.

    Journal: Infection and Immunity

    Article Title: Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

    doi:

    Figure Lengend Snippet: Immunoblot showing OmpL1 expression at different IPTG concentrations. A culture of E. coli XL1-Blue containing plasmid pMMB66-OmpL1 was grown to log phase, separated into four equal volumes, and treated with various concentrations of IPTG. Cells were incubated for an additional 3 h in a shaker incubator at 37°C, pelleted in a microcentrifuge, resuspended in final sample buffer, and analyzed by SDS-PAGE and immunoblotting. A companion SDS-polyacrylamide gel loaded with the same four samples was stained with Coomassie brilliant blue to verify that all four lanes contained comparable amounts of material (data not shown). The locations of molecular size standards are shown (in kilodaltons) on the left. Even though use of the tac promoter typically results in relatively high background expression levels, expression using plasmid pMMB66-OmpL1 was not detectable without IPTG induction.

    Article Snippet: Expression of OmpL1-M was studied in assays using a variety of different host strains, and E. coli XL1-Blue was found to produce the most consistent results.

    Techniques: Expressing, Plasmid Preparation, Incubation, SDS Page, Staining

    Coomassie brilliant blue-stained SDS-polyacrylamide gel showing material representative of E. coli membrane fractions used to immunize hamsters. Lanes: 1 and 2, E. coli XL1-Blue with plasmids pMMB66 and pMMB66-OmpL1, respectively; 3 and 4, E. coli JM109(DE3) with plasmids pET15b and pET15b-LipL41*, respectively. Locations of the OmpL1 doublet in lane 2 and of LipL41 in lane 4 are shown (arrowheads). Positions of molecular size markers (M) are given in kilodaltons.

    Journal: Infection and Immunity

    Article Title: Leptospiral Outer Membrane Proteins OmpL1 and LipL41 Exhibit Synergistic Immunoprotection

    doi:

    Figure Lengend Snippet: Coomassie brilliant blue-stained SDS-polyacrylamide gel showing material representative of E. coli membrane fractions used to immunize hamsters. Lanes: 1 and 2, E. coli XL1-Blue with plasmids pMMB66 and pMMB66-OmpL1, respectively; 3 and 4, E. coli JM109(DE3) with plasmids pET15b and pET15b-LipL41*, respectively. Locations of the OmpL1 doublet in lane 2 and of LipL41 in lane 4 are shown (arrowheads). Positions of molecular size markers (M) are given in kilodaltons.

    Article Snippet: Expression of OmpL1-M was studied in assays using a variety of different host strains, and E. coli XL1-Blue was found to produce the most consistent results.

    Techniques: Staining

    SDS gel electrophoretic analysis of cell lysates and purified enzyme fractions. (A) Native FB188 HDAH purified from Bordetella/Alcaligenes strain FB188 (DSM 11172). Lane 1, protein marker (purified FB188 HDAH prior to amino acid sequence analysis). (B) FB188 HDAH purified by IMAC from E. coli BL21 harboring pQEB-AH-NHis. A benchmark prestained protein ladder (Gibco-Life Science, Karlsruhe, Germany) (lane 1), crude cell extract (lane 2), and purified protein (lane 3) were fractionated on an SDS-10% polyacrylamide gel. Protein bands were visualized by Coomassie staining. (C) Western blotting using the anti-AHFB188 antibody (lanes are as described for panel B). (D) Western blot analysis of FB188 crude cell extract with anti-AHFB188 antibody. Lane 1, benchmark prestained protein ladder; lane 2, purified recombinant FB188 HDAH; lane 3, FB188 cell extract; lane 4, E. coli XL1-Blue cell extract.

    Journal: Journal of Bacteriology

    Article Title: A New Amidohydrolase from Bordetella or Alcaligenes Strain FB188 with Similarities to Histone Deacetylases

    doi: 10.1128/JB.186.8.2328-2339.2004

    Figure Lengend Snippet: SDS gel electrophoretic analysis of cell lysates and purified enzyme fractions. (A) Native FB188 HDAH purified from Bordetella/Alcaligenes strain FB188 (DSM 11172). Lane 1, protein marker (purified FB188 HDAH prior to amino acid sequence analysis). (B) FB188 HDAH purified by IMAC from E. coli BL21 harboring pQEB-AH-NHis. A benchmark prestained protein ladder (Gibco-Life Science, Karlsruhe, Germany) (lane 1), crude cell extract (lane 2), and purified protein (lane 3) were fractionated on an SDS-10% polyacrylamide gel. Protein bands were visualized by Coomassie staining. (C) Western blotting using the anti-AHFB188 antibody (lanes are as described for panel B). (D) Western blot analysis of FB188 crude cell extract with anti-AHFB188 antibody. Lane 1, benchmark prestained protein ladder; lane 2, purified recombinant FB188 HDAH; lane 3, FB188 cell extract; lane 4, E. coli XL1-Blue cell extract.

    Article Snippet: E. coli strains XL1-Blue and BL21 were purchased from Stratagene (La Jolla, Calif.).

    Techniques: SDS-Gel, Purification, Marker, Sequencing, Staining, Western Blot, Recombinant