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SDS-PAGE analysis of purified protein recombinant SllB in E. coli <t>BL21</t> cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.
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Images

1) Product Images from "Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification"

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2012.890

SDS-PAGE analysis of purified protein recombinant SllB in E. coli BL21 cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.
Figure Legend Snippet: SDS-PAGE analysis of purified protein recombinant SllB in E. coli BL21 cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.

Techniques Used: SDS Page, Purification, Recombinant, Marker

A transmission electron microscopic observation on the E. coli BL21 with recombinant protein. (A) Normal E. coli BL21 was treated as control. (B) The E. coli BL21 cells recombinant S-layer protein. (C) Crystal lattice structures on surface of the E. coli BL21 cells recombinant S-layer protein.
Figure Legend Snippet: A transmission electron microscopic observation on the E. coli BL21 with recombinant protein. (A) Normal E. coli BL21 was treated as control. (B) The E. coli BL21 cells recombinant S-layer protein. (C) Crystal lattice structures on surface of the E. coli BL21 cells recombinant S-layer protein.

Techniques Used: Transmission Assay, Recombinant

Expression of sllB in E. coli BL21 cells. (A) SDS-PAGE analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; lane M represent Takara Protein Marker (Broad). (B) Western blot analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; M, Takara Protein Marker (Broad). Lane M1, Precision plus protein standards; lane M2, perfect protein marker. Note the band pointed with arrows is the recombinant S-layer protein.
Figure Legend Snippet: Expression of sllB in E. coli BL21 cells. (A) SDS-PAGE analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; lane M represent Takara Protein Marker (Broad). (B) Western blot analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; M, Takara Protein Marker (Broad). Lane M1, Precision plus protein standards; lane M2, perfect protein marker. Note the band pointed with arrows is the recombinant S-layer protein.

Techniques Used: Expressing, SDS Page, Marker, Western Blot, Recombinant

2) Product Images from "Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine"

Article Title: Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine

Journal: Scientific Reports

doi: 10.1038/srep39235

HPTLC chromatogram of total lipid extracts from phage isolates. Lane 1: Escherichia coli BL21 positive control, lane 2: wild-type phage T7; lane 3: mutant phage T7::PhoE. Arrow head indicates the lipid species that is particularly enriched in the T7::PhoE mutant.
Figure Legend Snippet: HPTLC chromatogram of total lipid extracts from phage isolates. Lane 1: Escherichia coli BL21 positive control, lane 2: wild-type phage T7; lane 3: mutant phage T7::PhoE. Arrow head indicates the lipid species that is particularly enriched in the T7::PhoE mutant.

Techniques Used: High Performance Thin Layer Chromatography, Positive Control, Mutagenesis

3) Product Images from "Lysis Delay and Burst Shrinkage of Coliphage T7 by Deletion of Terminator Tφ Reversed by Deletion of Early Genes"

Article Title: Lysis Delay and Burst Shrinkage of Coliphage T7 by Deletion of Terminator Tφ Reversed by Deletion of Early Genes

Journal: Journal of Virology

doi: 10.1128/JVI.03274-13

Effects of holin overproduction on burst size and lysis time of T7 phage infection. (A and B) The T7 phage burst size (A) and lysis time (B) were measured with infection of the BL21 host carrying a pET21 vector containing the holin gene under 1.5 mM IPTG-induced
Figure Legend Snippet: Effects of holin overproduction on burst size and lysis time of T7 phage infection. (A and B) The T7 phage burst size (A) and lysis time (B) were measured with infection of the BL21 host carrying a pET21 vector containing the holin gene under 1.5 mM IPTG-induced

Techniques Used: Lysis, Infection, Plasmid Preparation

Related Articles

Clone Assay:

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Centrifugation:

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene). .. The E. coli cells were pelleted by centrifugation, resuspended in PBS buffer (200 μl/tube), and ultrasonicly disrupted.

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Amplification:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
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Filtration:

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Mass Spectrometry:

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Synthesized:

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
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Construct:

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
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Incubation:

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Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
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Stripping Membranes:

Article Title: Bacteriophage biodistribution and infectivity from honeybee to bee larvae using a T7 phage model
Article Snippet: E. coli BL21 (Stratagene) was the strain used as T7, T1 and T4 phage propagation strain. .. For phage propagation, 5 μL of phage suspension were spread evenly on host bacterial lawns using a paper strip and incubated O/N at 37 °C.

Infection:

Article Title: Lysis Delay and Burst Shrinkage of Coliphage T7 by Deletion of Terminator Tφ Reversed by Deletion of Early Genes
Article Snippet: .. E. coli BL21 (Stratagene) was used as the host organism for T7 phage preparation and infection. .. T7 phage was purchased from the ATCC (BAA-1025-B2).

Expressing:

Article Title: Tuning the Mycobacterium tuberculosis Alternative Sigma Factor SigF through the Multidomain Regulator Rv1364c and Osmosensory Kinase Protein Kinase D
Article Snippet: .. E. coli DH5α (Novagen) was used as a host strain for cloning purposes, and E. coli BL21(DE3) (Stratagene) was used as a host strain for the expression of recombinant proteins. .. E. coli cells were grown at 37°C in Luria-Bertani (LB) broth or LB agar (Difco) plates supplemented with 100 μg/ml of ampicillin and/or 25 μg/ml of kanamycin, when needed.

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: Paragraph title: Expression, western blotting and purification of sslB in E. coli BL21 (DE3) ... 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: The untagged merlin-1 tail domain (residues 503–595) was amplified and cloned into pET24b expression vector (Novagen) using the Nde I and Xho I restriction sites. .. Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail).

Article Title: Microbial Synthesis of Non-Natural Anthraquinone Glucosides Displaying Superior Antiproliferative Properties
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Article Title: The adaptor protein PID1 regulates receptor-dependent endocytosis of postprandial triglyceride-rich lipoproteins
Article Snippet: .. 2.6 GST-pulldown experiments For purification of GST-fusion proteins, the GST expression vector (pGex-6P, GE Healthcare) as well as the Flag-expression vector pCMV-Tag2B, both containing the respective insert (murine PID1 ; wild type and mutated LRP1 ICD; see for structure and mutations of the intracellular LRP1 domain) were transformed into E. coli BL21 (Stratagene). .. GST fusion proteins were purified via Glutathion Sepharose 4B (GE Healthcare).

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ). .. The vectors can also be cotransformed in E. coli Rosetta 2 (DE3) (Novagen), which contain rare codons that are seldom used in E. coli ( ).

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
Article Snippet: .. Individual CD155 expression constructs were transformed and expressed under control of the T7 promoter in Escherichia coli BL21 (λ DE3) (Stratagene, La Jolla, California). .. The bacteria were harvested by centrifugation at 8000 g (10 minutes, 4°C) and the resulting pellets were resuspended in extraction buffer (20 mM Tris-HCl, pH 7.5, 100 mM NaCl) containing lysozyme (Sigma-Aldrich, St Louis, Missouri).

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: After Nde I and Hind III digestion, the scFv fragment was inserted into pRB199 bacterial expression vector that had been engineered with the sequence for domains II and III of Pseudomonas exotoxin A (PE38KDEL) according to a previously described protocol. .. The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: .. 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Eight peptides at a length of approximately 30 amino acids and an overlap of 10 amino acids spanning the complete sequence of preS (genotype A, subtype adw2; Supplemental ) were synthesized by a Fmoc (9-fluorenylmethoxycarbonyl) - strategy with HBTU [2-(1H-Benzotriazol-1-yl)1,1,3,3 tetramethyluronium hexafluorophosphat] activation (Liberty Microwave Peptide Synthesis, CEM Corporation, Matthews, NC) as described previously ( ).

Western Blot:

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: Paragraph title: Expression, western blotting and purification of sslB in E. coli BL21 (DE3) ... 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

Transformation Assay:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: .. The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. At A 600 = 0.6, isopropyl-β- d -thio-galactopyranoside was added to a final concentration of 1 m m .

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: .. The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene). ..

Article Title: The adaptor protein PID1 regulates receptor-dependent endocytosis of postprandial triglyceride-rich lipoproteins
Article Snippet: .. 2.6 GST-pulldown experiments For purification of GST-fusion proteins, the GST expression vector (pGex-6P, GE Healthcare) as well as the Flag-expression vector pCMV-Tag2B, both containing the respective insert (murine PID1 ; wild type and mutated LRP1 ICD; see for structure and mutations of the intracellular LRP1 domain) were transformed into E. coli BL21 (Stratagene). .. GST fusion proteins were purified via Glutathion Sepharose 4B (GE Healthcare).

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: The plasmids are then transformed into host competent cells— E. coli XL1 Blue or E. coli TOP10 (Invitrogen)—isolated and sequenced. .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ).

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
Article Snippet: .. Individual CD155 expression constructs were transformed and expressed under control of the T7 promoter in Escherichia coli BL21 (λ DE3) (Stratagene, La Jolla, California). .. The bacteria were harvested by centrifugation at 8000 g (10 minutes, 4°C) and the resulting pellets were resuspended in extraction buffer (20 mM Tris-HCl, pH 7.5, 100 mM NaCl) containing lysozyme (Sigma-Aldrich, St Louis, Missouri).

Over Expression:

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: Paragraph title: Gene cloning, gene overexpression, and recombinant protein purification ... The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene).

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: The pET cloning system uses a strong T7 promoter, facilitating a robust overexpression of the protein encoded by the cloned gene. .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ).

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: .. E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments. .. E. coli BW25113 Δ bamB Δ tolC is an antibiotic hypersensitive , and was used for susceptibility testing of Mphs cloning into pGDP4.

High Performance Liquid Chromatography:

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Peptides were purified by preparative HPLC and their identity was confirmed by mass spectrometry (Microflex MALDI-TOF, Bruker, Billerica, MA).

Chromatography:

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail). .. Cells were pooled and lysed in 50 m M Tris, 300 m M NaCl (pH 8), and complete mini protease inhibitor tablet (Roche) and ultracentrifuged at 95,834 g for 1 h. Proteins were copurified using a GST FF chromatography affinity column (GE Life Sciences) and eluted with 10 m M reduced glutathione.

Concentration Assay:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. At A 600 = 0.6, isopropyl-β- d -thio-galactopyranoside was added to a final concentration of 1 m m .

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene). .. Isopropyl-β-D-thiogalactopyranoside (IPTG) was then added (50 μl of a 100 mM stock, final concentration 1 mM) for 3 h to induce the tac promoter.

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments. .. Antibiotic susceptibility testing was performed according to the Clinical and Laboratory Standards Institute guidelines for determining minimal inhibitory concentration (MIC) in 96-well plates (Sarstedt, Germany) in duplicate.

Protease Inhibitor:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. Three hours after induction, cells were harvested and resuspended in tyrosine kinase induction medium containing 53 μ m indoleacrylic acid and grown for 2 h. Cells were harvested and resuspended in wash buffer (50 m m NaPO4 , 300 m m NaCl, 20 m m imidazole, pH 8.0) supplemented with Complete protease inhibitor, EDTA-free (Roche Applied Science) and lysed with a French press.

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail). .. Cells were pooled and lysed in 50 m M Tris, 300 m M NaCl (pH 8), and complete mini protease inhibitor tablet (Roche) and ultracentrifuged at 95,834 g for 1 h. Proteins were copurified using a GST FF chromatography affinity column (GE Life Sciences) and eluted with 10 m M reduced glutathione.

Cell Culture:

Article Title: Bacteriophage biodistribution and infectivity from honeybee to bee larvae using a T7 phage model
Article Snippet: E. coli BL21 (Stratagene) was the strain used as T7, T1 and T4 phage propagation strain. .. Bacteria were cultured at 37 °C overnight (O/N) in Tryptic Soy Broth (TSB, VWR) or Tryptic Soy Agar medium (TSA; TSB containing 1.5% (w/v) agar, NZYTech).

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: E. coli was routinely cultured on LB-Lennox (Bioshop) at 37 °C overnight, and either 100 µg mL−1 ampicillin or 50 µg mL−1 kanamycin was used for plasmid selection. .. E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments.

Generated:

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: The DmAb14-scFv IT was generated by PCR using parental DmAb14-scFv plasmid as a template and primers introduced at the Nde I and Hind III restriction enzyme sites. .. The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Polymerase Chain Reaction:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: A synthetic version of the STAT3 gene, codon-optimized for Escherichia coli (DNA 2.0) encoding amino acids 127–722, was amplified by PCR and ligated into pETm11-SUMO3 (European Molecular Biology Laboratory (EMBL)). .. The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid.

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: The DmAb14-scFv IT was generated by PCR using parental DmAb14-scFv plasmid as a template and primers introduced at the Nde I and Hind III restriction enzyme sites. .. The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Sonication:

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene). .. Cells were broken by sonication, and the recombinant protein was purified using a nickel–nitrilotriacetic acid preparatory column (Qiagene) following the manufacturer’s protocol ( ).

Recombinant:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: Paragraph title: Recombinant pSTAT3 Protein ... The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid.

Article Title: Tuning the Mycobacterium tuberculosis Alternative Sigma Factor SigF through the Multidomain Regulator Rv1364c and Osmosensory Kinase Protein Kinase D
Article Snippet: .. E. coli DH5α (Novagen) was used as a host strain for cloning purposes, and E. coli BL21(DE3) (Stratagene) was used as a host strain for the expression of recombinant proteins. .. E. coli cells were grown at 37°C in Luria-Bertani (LB) broth or LB agar (Difco) plates supplemented with 100 μg/ml of ampicillin and/or 25 μg/ml of kanamycin, when needed.

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: Paragraph title: Gene cloning, gene overexpression, and recombinant protein purification ... The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene).

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: Paragraph title: Preparation of recombinant immunotoxins ... The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: .. 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Eight peptides at a length of approximately 30 amino acids and an overlap of 10 amino acids spanning the complete sequence of preS (genotype A, subtype adw2; Supplemental ) were synthesized by a Fmoc (9-fluorenylmethoxycarbonyl) - strategy with HBTU [2-(1H-Benzotriazol-1-yl)1,1,3,3 tetramethyluronium hexafluorophosphat] activation (Liberty Microwave Peptide Synthesis, CEM Corporation, Matthews, NC) as described previously ( ).

Mutagenesis:

Article Title: Lysis Delay and Burst Shrinkage of Coliphage T7 by Deletion of Terminator Tφ Reversed by Deletion of Early Genes
Article Snippet: E. coli BL21 (Stratagene) was used as the host organism for T7 phage preparation and infection. .. A Tφ-lacking T7 phage mutant was constructed with a deletion of 31 bp of the Tφ terminator.

Size-exclusion Chromatography:

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene). .. The IT proteins were then reduced, refolded, and further purified as monomers (67 kDa) by using ion-exchange and size-exclusion chromatography to greater than 90% purity.

Purification:

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene). .. Cells were broken by sonication, and the recombinant protein was purified using a nickel–nitrilotriacetic acid preparatory column (Qiagene) following the manufacturer’s protocol ( ).

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: Paragraph title: Expression, western blotting and purification of sslB in E. coli BL21 (DE3) ... 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail). .. The head:tail complex was further purified using a Superdex 75 26/60 gel filtration chromatography column (GE Life Sciences) equilibrated with 50 m M Tris and 300 m M NaCl (pH 8).

Article Title: The adaptor protein PID1 regulates receptor-dependent endocytosis of postprandial triglyceride-rich lipoproteins
Article Snippet: .. 2.6 GST-pulldown experiments For purification of GST-fusion proteins, the GST expression vector (pGex-6P, GE Healthcare) as well as the Flag-expression vector pCMV-Tag2B, both containing the respective insert (murine PID1 ; wild type and mutated LRP1 ICD; see for structure and mutations of the intracellular LRP1 domain) were transformed into E. coli BL21 (Stratagene). .. GST fusion proteins were purified via Glutathion Sepharose 4B (GE Healthcare).

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: PqqE has been expressed and purified as a His6 -tag construct (either N-terminal or C-terminal), thus the vectors pET28 (Novagen) have been used for cloning purposes ( ; ; ). .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ).

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
Article Snippet: Paragraph title: Construction, Expression, and Purification of CD155-Derived Polypeptides ... Individual CD155 expression constructs were transformed and expressed under control of the T7 promoter in Escherichia coli BL21 (λ DE3) (Stratagene, La Jolla, California).

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene). .. The IT proteins were then reduced, refolded, and further purified as monomers (67 kDa) by using ion-exchange and size-exclusion chromatography to greater than 90% purity.

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: .. 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Eight peptides at a length of approximately 30 amino acids and an overlap of 10 amino acids spanning the complete sequence of preS (genotype A, subtype adw2; Supplemental ) were synthesized by a Fmoc (9-fluorenylmethoxycarbonyl) - strategy with HBTU [2-(1H-Benzotriazol-1-yl)1,1,3,3 tetramethyluronium hexafluorophosphat] activation (Liberty Microwave Peptide Synthesis, CEM Corporation, Matthews, NC) as described previously ( ).

Protein Purification:

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: Paragraph title: Gene cloning, gene overexpression, and recombinant protein purification ... The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene).

Sequencing:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: .. The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. At A 600 = 0.6, isopropyl-β- d -thio-galactopyranoside was added to a final concentration of 1 m m .

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ). .. The vectors can also be cotransformed in E. coli Rosetta 2 (DE3) (Novagen), which contain rare codons that are seldom used in E. coli ( ).

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: After Nde I and Hind III digestion, the scFv fragment was inserted into pRB199 bacterial expression vector that had been engineered with the sequence for domains II and III of Pseudomonas exotoxin A (PE38KDEL) according to a previously described protocol. .. The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: The codon optimized mphB sequence is in Supplementary Table . mphI was previously cloned into pET22b . pGDP4 is a derivative of pET28a with the multiple-cloning site under the control of the constitutive lac promoter . mphI was subcloned into pGDP4 using the XbaI and XhoI restriction sites. .. E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments.

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: .. 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Eight peptides at a length of approximately 30 amino acids and an overlap of 10 amino acids spanning the complete sequence of preS (genotype A, subtype adw2; Supplemental ) were synthesized by a Fmoc (9-fluorenylmethoxycarbonyl) - strategy with HBTU [2-(1H-Benzotriazol-1-yl)1,1,3,3 tetramethyluronium hexafluorophosphat] activation (Liberty Microwave Peptide Synthesis, CEM Corporation, Matthews, NC) as described previously ( ).

Affinity Chromatography:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. The lysate was ultracentrifuged (180,000 × g , 60 min, 4 °C), and the supernatant was used for affinity chromatography on a 1-ml HisTrap HP column (GE Healthcare).

Affinity Column:

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail). .. Cells were pooled and lysed in 50 m M Tris, 300 m M NaCl (pH 8), and complete mini protease inhibitor tablet (Roche) and ultracentrifuged at 95,834 g for 1 h. Proteins were copurified using a GST FF chromatography affinity column (GE Life Sciences) and eluted with 10 m M reduced glutathione.

Plasmid Preparation:

Article Title: Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells *Galiellalactone Is a Direct Inhibitor of the Transcription Factor STAT3 in Prostate Cancer Cells * ♦
Article Snippet: .. The plasmid was verified by sequencing and transformed into E. coli BL21(DE3) TKB1 (Stratagene), which harbors a plasmid-encoded tyrosine kinase gene behind a promoter inducible with indoleacrylic acid. .. At A 600 = 0.6, isopropyl-β- d -thio-galactopyranoside was added to a final concentration of 1 m m .

Article Title: Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine
Article Snippet: The phage host was E. coli BL21 (Stratagene). .. Plasmid pKD46, an ampicillin-resistant and temperature sensitive plasmid that encodes the lambda Red genes (exo, beta and gam ) was used to prepare recombineering competent cells.

Article Title: Dramatic performance of Clostridium thermocellum explained by its wide range of cellulase modalities
Article Snippet: .. The plasmid with gene insertion was then transformed into Escherichia coli BL21(DE3) (Stratagene). ..

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: Expression, western blotting and purification of sslB in E. coli BL21 (DE3) A pET28a(+)-sllB plasmid was prepared using the MiniBEST Plasmid Purification kit ver. .. 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

Article Title: Unfurling of the band 4.1, ezrin, radixin, moesin (FERM) domain of the merlin tumor suppressor
Article Snippet: The untagged merlin-1 tail domain (residues 503–595) was amplified and cloned into pET24b expression vector (Novagen) using the Nde I and Xho I restriction sites. .. Proteins were expressed in Escherichia coli BL21(DE3)RIL (Stratagene) at 25°C for 20 h in Luria–Bertani medium with ampicillin (GST-head) or kanamycin (tail).

Article Title: The adaptor protein PID1 regulates receptor-dependent endocytosis of postprandial triglyceride-rich lipoproteins
Article Snippet: .. 2.6 GST-pulldown experiments For purification of GST-fusion proteins, the GST expression vector (pGex-6P, GE Healthcare) as well as the Flag-expression vector pCMV-Tag2B, both containing the respective insert (murine PID1 ; wild type and mutated LRP1 ICD; see for structure and mutations of the intracellular LRP1 domain) were transformed into E. coli BL21 (Stratagene). .. GST fusion proteins were purified via Glutathion Sepharose 4B (GE Healthcare).

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ). .. The vectors can also be cotransformed in E. coli Rosetta 2 (DE3) (Novagen), which contain rare codons that are seldom used in E. coli ( ).

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
Article Snippet: Complementary DNAs encoding the CD155 D2 (aa 145–237) or sub-D2 (aa 145–199) were synthesized (GenScript, Piscataway, New Jersey) for insertion into the Eco RI/ Hin DIII sites of the pET43.1a(+) expression vector (EMD Biosciences, San Diego, California) and the sequences verified (GenScript). .. Individual CD155 expression constructs were transformed and expressed under control of the T7 promoter in Escherichia coli BL21 (λ DE3) (Stratagene, La Jolla, California).

Article Title: Affinity-matured recombinant immunotoxin targeting gangliosides 3′-isoLM1 and 3′,6'-isoLD1 on malignant gliomas
Article Snippet: After Nde I and Hind III digestion, the scFv fragment was inserted into pRB199 bacterial expression vector that had been engineered with the sequence for domains II and III of Pseudomonas exotoxin A (PE38KDEL) according to a previously described protocol. .. The parental DmAb14-scFv IT was expressed under control of the T7 promoter in E. coli BL21 (λ DE3) (Stratagene).

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: E. coli was routinely cultured on LB-Lennox (Bioshop) at 37 °C overnight, and either 100 µg mL−1 ampicillin or 50 µg mL−1 kanamycin was used for plasmid selection. .. E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments.

Selection:

Article Title: Microbial Synthesis of Non-Natural Anthraquinone Glucosides Displaying Superior Antiproliferative Properties
Article Snippet: E. coli BL21 (DE3, Stratagene, La Jolla, CA, USA) was used as expression and biotransformation hosts. .. Luria-Bertani (LB) plates and broth media supplemented with an appropriate antibiotic (kanamycin 50 μg/mL) was used for the E. coli growth, colony selection, culture preparation, and biotransformation.

Article Title: The evolution of substrate discrimination in macrolide antibiotic resistance enzymes
Article Snippet: E. coli was routinely cultured on LB-Lennox (Bioshop) at 37 °C overnight, and either 100 µg mL−1 ampicillin or 50 µg mL−1 kanamycin was used for plasmid selection. .. E. coli BL21(DE3) (Stratagene, USA)was used for protein overexpression experiments.

Activation Assay:

Article Title: Immunotherapy With the PreS-based Grass Pollen Allergy Vaccine BM32 Induces Antibody Responses Protecting Against Hepatitis B Infection
Article Snippet: 2.1 Expression and Purification of Recombinant PreS, Synthesis of PreS Overlapping Peptides, Sequence Alignments The procedure of the expression and purification of a hexahistidine-tagged recombinant preS protein (preS1 + preS2; genotype A; subtype adw2, GenBank: AAT28735.1 ) in Escherichia coli BL21 (DE3, Stratagene, La Jolla, CA) is described elsewhere ( ). .. Eight peptides at a length of approximately 30 amino acids and an overlap of 10 amino acids spanning the complete sequence of preS (genotype A, subtype adw2; Supplemental ) were synthesized by a Fmoc (9-fluorenylmethoxycarbonyl) - strategy with HBTU [2-(1H-Benzotriazol-1-yl)1,1,3,3 tetramethyluronium hexafluorophosphat] activation (Liberty Microwave Peptide Synthesis, CEM Corporation, Matthews, NC) as described previously ( ).

Phage Preparation:

Article Title: Lysis Delay and Burst Shrinkage of Coliphage T7 by Deletion of Terminator Tφ Reversed by Deletion of Early Genes
Article Snippet: .. E. coli BL21 (Stratagene) was used as the host organism for T7 phage preparation and infection. .. T7 phage was purchased from the ATCC (BAA-1025-B2).

Lysis:

Article Title: Validation of an Immunohistochemistry Assay for Detection of CD155, the Poliovirus Receptor, in Malignant Gliomas
Article Snippet: Individual CD155 expression constructs were transformed and expressed under control of the T7 promoter in Escherichia coli BL21 (λ DE3) (Stratagene, La Jolla, California). .. The bacterial suspension was incubated at room temperature (20 minutes) with gentle shaking for complete cell lysis to occur.

Positron Emission Tomography:

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: The pET cloning system uses a strong T7 promoter, facilitating a robust overexpression of the protein encoded by the cloned gene. .. Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ).

Chick Chorioallantoic Membrane Assay:

Article Title: Methods for Expression, Purification, and Characterization of PqqE, a Radical SAM Enzyme in the PQQ Biosynthetic Pathway
Article Snippet: Once the sequence is confirmed, the plasmid harboring the correct construct is cotransformed into E. coli BL21(DE3) (Stratagene) with vectors expressing the E. coli suf ABCDSE operon (pPH151) genes, which encodes proteins involved in Fe–S cluster repair and assembly ( ). .. The pET vector contains a kanamycin (Kan) resistance gene, while the pPH151 is resistant to chloramphenicol (Cam); thus, the transformed strain is Kan and Cam resistant and can be grown in selective media with both antibiotics.

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    Stratagene bl21 codon plus de3 ril
    Western blot analysis of DsbA:HIV-1Pr expression on total cell extracts of <t>BL21-Codon</t> <t>Plus-(DE3)-RIL</t> E. coli cells containing the pET39-DsbA:HIV-1Pr plasmid and using different cultivation media. (Indicated below each panel): the cells were collected at different times after adding 1 mM IPTG (0, 0.5, 1, or 3 hours, as indicated above each lane). Protein expression induced at A) the early-exponential phase, B) the middle-exponential phase, and C) the stationary phase. An amount of cells corresponding to 1 mL (panel A) or 0.5 mL (panels B and C) of culture was loaded in each lane. D) Comparison of DsbA:HIV-1Pr production level obtained using different growth conditions. The first bar represents the best conditions identified using different E. coli strains (see Figure 1). In all cases variability among replicate cultures was lower than 10%. DsbA:HIV-1Pr expression was detected using anti-His-tag-specific antibodies. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.
    Bl21 Codon Plus De3 Ril, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Stratagene ptd drbd expression utilized bl21 codon
    <t>PTD-DRBD</t> siRNA delivery into T cells and HUVECs ( a ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics of dividing Jurkat dGFP cells following treatment with GFP2 siRNA plus PTD-DRBD, Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2), as indicated. ( b ) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 12 h post-treatment of GAPDH siRNA or GFP2 (Con) siRNA plus PTD-DRBD, GAPDH siRNA plus Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2) in Jurkat cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. ( c ) Flow cytometry histogram analysis of PTD-DRBD mediated CD4 or CD8 RNAi response at 1 day post-treatment of mouse primary T cells, as indicated. ( d ) Quantitative RT-PCR analysis of endogenous CD4, CD8 or CD90 mRNA expression at 12 and 24 h post-treatment of PTD-DRBD CD4 or CD8 siRNAs in primary T cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock control. *(P
    Ptd Drbd Expression Utilized Bl21 Codon, supplied by Stratagene, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene e coli protease deficient expression strain bl21
    Competitive binding of cannabinoid ligands on <t>BL21-107</t> membranes expressing CB2. The assay was performed in triplicate as described in Materials and Methods.
    E Coli Protease Deficient Expression Strain Bl21, supplied by Stratagene, used in various techniques. Bioz Stars score: 70/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Stratagene e coli bl21
    SDS-PAGE analysis of purified protein recombinant SllB in E. coli <t>BL21</t> cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.
    E Coli Bl21, supplied by Stratagene, used in various techniques. Bioz Stars score: 96/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Western blot analysis of DsbA:HIV-1Pr expression on total cell extracts of BL21-Codon Plus-(DE3)-RIL E. coli cells containing the pET39-DsbA:HIV-1Pr plasmid and using different cultivation media. (Indicated below each panel): the cells were collected at different times after adding 1 mM IPTG (0, 0.5, 1, or 3 hours, as indicated above each lane). Protein expression induced at A) the early-exponential phase, B) the middle-exponential phase, and C) the stationary phase. An amount of cells corresponding to 1 mL (panel A) or 0.5 mL (panels B and C) of culture was loaded in each lane. D) Comparison of DsbA:HIV-1Pr production level obtained using different growth conditions. The first bar represents the best conditions identified using different E. coli strains (see Figure 1). In all cases variability among replicate cultures was lower than 10%. DsbA:HIV-1Pr expression was detected using anti-His-tag-specific antibodies. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Journal: Microbial Cell Factories

    Article Title: Optimizing HIV-1 protease production in Escherichia coli as fusion protein

    doi: 10.1186/1475-2859-10-53

    Figure Lengend Snippet: Western blot analysis of DsbA:HIV-1Pr expression on total cell extracts of BL21-Codon Plus-(DE3)-RIL E. coli cells containing the pET39-DsbA:HIV-1Pr plasmid and using different cultivation media. (Indicated below each panel): the cells were collected at different times after adding 1 mM IPTG (0, 0.5, 1, or 3 hours, as indicated above each lane). Protein expression induced at A) the early-exponential phase, B) the middle-exponential phase, and C) the stationary phase. An amount of cells corresponding to 1 mL (panel A) or 0.5 mL (panels B and C) of culture was loaded in each lane. D) Comparison of DsbA:HIV-1Pr production level obtained using different growth conditions. The first bar represents the best conditions identified using different E. coli strains (see Figure 1). In all cases variability among replicate cultures was lower than 10%. DsbA:HIV-1Pr expression was detected using anti-His-tag-specific antibodies. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Article Snippet: Strain, growth conditions, and enzyme expression For protein expression, plasmids containing the HIV-1Pr cDNA were transferred into different E. coli strains: BL21(DE3)pLysS (Novagen, Madison, Wisconsin, USA), BL21-Codon Plus(DE3)-RIL (Stratagene, La Jolla, California, USA), BL21-Star(DE3) (Invitrogen, Carslbad, California, USA), KRX (Promega, Madison, Wisconsin, USA), and C41(DE3), C41(DE3)pLysS, C43(DE3), C43(DE3)pLysS (Lucigen, Middleton, Wisconsin, USA) host cells.

    Techniques: Western Blot, Expressing, Plasmid Preparation, Recombinant

    Western blot analysis of DsbA:HIV-1Pr protein expression on total cell extracts of different E. coli strains containing the pET39-DsbA:HIV-1Pr plasmid. As detected using anti-His-tag-specific antibodies. Cells were grown in LB medium at 37°C, protein expression was induced using 1 mM IPTG at early-exponential phase, and cells were harvested after 1 hour. E. coli strains are: 1: BL21(DE3)pLysS; 2: C41(DE3); 3: C43(DE3); 4: C41(DE3)pLysS; 5: C43(DE3)pLysS; 6: KRX; 7: BL21-CodonPlus(DE3)-RIL; and 8: BL21(DE3)Star. In all cases variability among replicate cultures was lower than 10%. The amount of cells corresponding to 1 mL of culture was loaded in each lane. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Journal: Microbial Cell Factories

    Article Title: Optimizing HIV-1 protease production in Escherichia coli as fusion protein

    doi: 10.1186/1475-2859-10-53

    Figure Lengend Snippet: Western blot analysis of DsbA:HIV-1Pr protein expression on total cell extracts of different E. coli strains containing the pET39-DsbA:HIV-1Pr plasmid. As detected using anti-His-tag-specific antibodies. Cells were grown in LB medium at 37°C, protein expression was induced using 1 mM IPTG at early-exponential phase, and cells were harvested after 1 hour. E. coli strains are: 1: BL21(DE3)pLysS; 2: C41(DE3); 3: C43(DE3); 4: C41(DE3)pLysS; 5: C43(DE3)pLysS; 6: KRX; 7: BL21-CodonPlus(DE3)-RIL; and 8: BL21(DE3)Star. In all cases variability among replicate cultures was lower than 10%. The amount of cells corresponding to 1 mL of culture was loaded in each lane. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Article Snippet: Strain, growth conditions, and enzyme expression For protein expression, plasmids containing the HIV-1Pr cDNA were transferred into different E. coli strains: BL21(DE3)pLysS (Novagen, Madison, Wisconsin, USA), BL21-Codon Plus(DE3)-RIL (Stratagene, La Jolla, California, USA), BL21-Star(DE3) (Invitrogen, Carslbad, California, USA), KRX (Promega, Madison, Wisconsin, USA), and C41(DE3), C41(DE3)pLysS, C43(DE3), C43(DE3)pLysS (Lucigen, Middleton, Wisconsin, USA) host cells.

    Techniques: Western Blot, Expressing, Plasmid Preparation, Recombinant

    Amount of soluble DsbA:HIV-1Pr and phosphate salts effect on chimeric protein production . A) Comparison by means of Western blot analysis of the amount of DsbA:HIV-1Pr protein expression in the soluble (S) and insoluble (I) cell fractions as obtained after disruption of BL21(DE3)pLysS E. coli cells (growth in TB medium, protein expression induced at the early-exponential phase) or of BL21-Codon Plus-(DE3)-RIL E. coli cells (protein expression induced at the early and middle-exponential phase of growth). B) Western blot analysis of DsbA:HIV-1Pr protein expression in total cell extracts of recombinant BL21-Codon Plus-(DE3)-RIL E. coli cells grown in the presence of a phosphate salts buffer system. LB: cells grown in LB medium (control); a: cells grown in LB medium added of 6.78 g/L of NaH 2 PO 4 and 3 g/L of KH 2 PO 4 (such as in M9 broth); b: cells grown in LB medium to which 9.4 g/L of K 2 HPO 4 and 2.2 g/L of KH 2 PO 4 were added (such as in TB broth). Protein expression was induced in all samples with 1 mM IPTG; cells were harvested at 1 or 3 hours after adding IPTG. An amount of cells corresponding to 0.2 mL of culture was loaded in each lane. C) Summary results of DsbA:HIV-1Pr production (see panel B): the expression level of cells grown on LB medium is reported as 100% (= 1.8 mg/L). Cells were collected at 1 hour (empty bars) and 3 hours (black bars) from IPTG addition. Western blot analysis was carried out by using anti-His-tag-specific antibodies. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Journal: Microbial Cell Factories

    Article Title: Optimizing HIV-1 protease production in Escherichia coli as fusion protein

    doi: 10.1186/1475-2859-10-53

    Figure Lengend Snippet: Amount of soluble DsbA:HIV-1Pr and phosphate salts effect on chimeric protein production . A) Comparison by means of Western blot analysis of the amount of DsbA:HIV-1Pr protein expression in the soluble (S) and insoluble (I) cell fractions as obtained after disruption of BL21(DE3)pLysS E. coli cells (growth in TB medium, protein expression induced at the early-exponential phase) or of BL21-Codon Plus-(DE3)-RIL E. coli cells (protein expression induced at the early and middle-exponential phase of growth). B) Western blot analysis of DsbA:HIV-1Pr protein expression in total cell extracts of recombinant BL21-Codon Plus-(DE3)-RIL E. coli cells grown in the presence of a phosphate salts buffer system. LB: cells grown in LB medium (control); a: cells grown in LB medium added of 6.78 g/L of NaH 2 PO 4 and 3 g/L of KH 2 PO 4 (such as in M9 broth); b: cells grown in LB medium to which 9.4 g/L of K 2 HPO 4 and 2.2 g/L of KH 2 PO 4 were added (such as in TB broth). Protein expression was induced in all samples with 1 mM IPTG; cells were harvested at 1 or 3 hours after adding IPTG. An amount of cells corresponding to 0.2 mL of culture was loaded in each lane. C) Summary results of DsbA:HIV-1Pr production (see panel B): the expression level of cells grown on LB medium is reported as 100% (= 1.8 mg/L). Cells were collected at 1 hour (empty bars) and 3 hours (black bars) from IPTG addition. Western blot analysis was carried out by using anti-His-tag-specific antibodies. St: 0.5 μg of His-tagged recombinant D-amino acid oxidase.

    Article Snippet: Strain, growth conditions, and enzyme expression For protein expression, plasmids containing the HIV-1Pr cDNA were transferred into different E. coli strains: BL21(DE3)pLysS (Novagen, Madison, Wisconsin, USA), BL21-Codon Plus(DE3)-RIL (Stratagene, La Jolla, California, USA), BL21-Star(DE3) (Invitrogen, Carslbad, California, USA), KRX (Promega, Madison, Wisconsin, USA), and C41(DE3), C41(DE3)pLysS, C43(DE3), C43(DE3)pLysS (Lucigen, Middleton, Wisconsin, USA) host cells.

    Techniques: Western Blot, Expressing, Recombinant

    Effect of temperature of growth after adding IPTG on the expression of DsbA:HIV-1Pr . Western blot analysis of soluble (S) and insoluble (I) cell extracts as obtained after disruption of recombinant BL21-Codon Plus-(DE3)-RIL E. coli cells expressing DsbA:HIV-1Pr fusion protein grown in TB (A) or M9 (B) media at different temperatures. Protein expression was induced by adding 1 mM IPTG at the early or middle-exponential phase of growth. Experimental conditions as in Figure 2. C) Summary results of soluble (empty bars) vs. insoluble (black bars) level of DsbA:HIV-1Pr expression in cells induced at the middle-exponential phase of growth and using TB or M9 media (see panels A and B).

    Journal: Microbial Cell Factories

    Article Title: Optimizing HIV-1 protease production in Escherichia coli as fusion protein

    doi: 10.1186/1475-2859-10-53

    Figure Lengend Snippet: Effect of temperature of growth after adding IPTG on the expression of DsbA:HIV-1Pr . Western blot analysis of soluble (S) and insoluble (I) cell extracts as obtained after disruption of recombinant BL21-Codon Plus-(DE3)-RIL E. coli cells expressing DsbA:HIV-1Pr fusion protein grown in TB (A) or M9 (B) media at different temperatures. Protein expression was induced by adding 1 mM IPTG at the early or middle-exponential phase of growth. Experimental conditions as in Figure 2. C) Summary results of soluble (empty bars) vs. insoluble (black bars) level of DsbA:HIV-1Pr expression in cells induced at the middle-exponential phase of growth and using TB or M9 media (see panels A and B).

    Article Snippet: Strain, growth conditions, and enzyme expression For protein expression, plasmids containing the HIV-1Pr cDNA were transferred into different E. coli strains: BL21(DE3)pLysS (Novagen, Madison, Wisconsin, USA), BL21-Codon Plus(DE3)-RIL (Stratagene, La Jolla, California, USA), BL21-Star(DE3) (Invitrogen, Carslbad, California, USA), KRX (Promega, Madison, Wisconsin, USA), and C41(DE3), C41(DE3)pLysS, C43(DE3), C43(DE3)pLysS (Lucigen, Middleton, Wisconsin, USA) host cells.

    Techniques: Expressing, Western Blot, Recombinant

    PTD-DRBD siRNA delivery into T cells and HUVECs ( a ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics of dividing Jurkat dGFP cells following treatment with GFP2 siRNA plus PTD-DRBD, Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2), as indicated. ( b ) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 12 h post-treatment of GAPDH siRNA or GFP2 (Con) siRNA plus PTD-DRBD, GAPDH siRNA plus Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2) in Jurkat cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. ( c ) Flow cytometry histogram analysis of PTD-DRBD mediated CD4 or CD8 RNAi response at 1 day post-treatment of mouse primary T cells, as indicated. ( d ) Quantitative RT-PCR analysis of endogenous CD4, CD8 or CD90 mRNA expression at 12 and 24 h post-treatment of PTD-DRBD CD4 or CD8 siRNAs in primary T cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock control. *(P

    Journal: Nature biotechnology

    Article Title: Efficient siRNA Delivery into Primary Cells by Peptide Transduction-dsRNA Binding Domain (PTD-DRBD) Fusion Protein

    doi: 10.1038/nbt.1541

    Figure Lengend Snippet: PTD-DRBD siRNA delivery into T cells and HUVECs ( a ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics of dividing Jurkat dGFP cells following treatment with GFP2 siRNA plus PTD-DRBD, Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2), as indicated. ( b ) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 12 h post-treatment of GAPDH siRNA or GFP2 (Con) siRNA plus PTD-DRBD, GAPDH siRNA plus Lipofection-2000 (Lipofection) or RNAiMAX (Lipofection 2) in Jurkat cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. ( c ) Flow cytometry histogram analysis of PTD-DRBD mediated CD4 or CD8 RNAi response at 1 day post-treatment of mouse primary T cells, as indicated. ( d ) Quantitative RT-PCR analysis of endogenous CD4, CD8 or CD90 mRNA expression at 12 and 24 h post-treatment of PTD-DRBD CD4 or CD8 siRNAs in primary T cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock control. *(P

    Article Snippet: PTD-DRBD expression utilized BL21 codon plus (DH3) E.coli (Strategene) cells were transformed with pPTD-DRBD, cultured at 37°C in LB, then at 25°C for 12 h after induction with 400 μM IPTG.

    Techniques: Flow Cytometry, Cytometry, Quantitative RT-PCR, Expressing

    PTD-DRBD mediated siRNA delivery ( a ) Hypothetical cartoon of PTD-DRBD bound to siRNA. DRBD Ribbon structure derived from Ryter and Schultze 13 ( b ) Normalized RNAi knockdown of dGFP and dDsRed by PTD-DRBD:siRNA (left panel) and lipofection (right panel), as indicated, in H1299 dGFP/dDsRed cells. Mean values were normalized to percent control. ( c,d ) Single cell flow cytometry histogram analysis of dGFP RNAi response at 1 and 2 days post-treatment of H1299 dGFP/dDsRed cells, as indicated. ( e ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics following a single siRNA treatment of dividing H1299 dGFP/dDsRed cells. ( f ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics following multiple siRNA treatments of H1299 dGFP cells, as indicated. Mean values are normalized to percent control. ( g,h ) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 6 and 12 h post-treatment in H1299 cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. **(P

    Journal: Nature biotechnology

    Article Title: Efficient siRNA Delivery into Primary Cells by Peptide Transduction-dsRNA Binding Domain (PTD-DRBD) Fusion Protein

    doi: 10.1038/nbt.1541

    Figure Lengend Snippet: PTD-DRBD mediated siRNA delivery ( a ) Hypothetical cartoon of PTD-DRBD bound to siRNA. DRBD Ribbon structure derived from Ryter and Schultze 13 ( b ) Normalized RNAi knockdown of dGFP and dDsRed by PTD-DRBD:siRNA (left panel) and lipofection (right panel), as indicated, in H1299 dGFP/dDsRed cells. Mean values were normalized to percent control. ( c,d ) Single cell flow cytometry histogram analysis of dGFP RNAi response at 1 and 2 days post-treatment of H1299 dGFP/dDsRed cells, as indicated. ( e ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics following a single siRNA treatment of dividing H1299 dGFP/dDsRed cells. ( f ) Flow cytometry analysis of dGFP RNAi knockdown decay kinetics following multiple siRNA treatments of H1299 dGFP cells, as indicated. Mean values are normalized to percent control. ( g,h ) Quantitative RT-PCR analysis of endogenous GAPDH mRNA expression at 6 and 12 h post-treatment in H1299 cells, as indicated. Mean values normalized to β2 microglobulin and reported as percent of mock GAPDH control. **(P

    Article Snippet: PTD-DRBD expression utilized BL21 codon plus (DH3) E.coli (Strategene) cells were transformed with pPTD-DRBD, cultured at 37°C in LB, then at 25°C for 12 h after induction with 400 μM IPTG.

    Techniques: Derivative Assay, Flow Cytometry, Cytometry, Quantitative RT-PCR, Expressing

    PTD-DRBD mediated RNAi responses ( a ) Fluorescent microscopy analysis of H9 hESCs constitutively expressing GFP treated with with PTD-DRBD delivered GFP2 siRNA at 2 days post-addition. Black line outlines hESC colony on mouse feeder cell background. ( b ) Oct4 immunoblot analysis in HUES9 hESCs treated with mock (PBS), PTD-DRBD delivered Oct4 or control siRNAs at 2 days post-addition. ( c ) Cell division curve of human HUES9 embryonic stem cells treated with mock (PBS), PTD-DRBD delivered Oct4 or control siRNAs, as indicated. ( d ) Immunohistochemistry analysis of Oct4 and SSEA4 expression in HUES9 hESCs at 2 days post-treatment with mock (PBS), PTD-DRB delivered Oct4 or control siRNAs. Anti-Oct4 antibodies (red), anti-SSEA-4 antibodies (green), genomic DNA (blue). ( e ) Immuno-histochemistry analysis of GATA6 and SSEA4 expression in HUES9 hESCs at 10 days post-treatment with mock (PBS), PTD-DRB delivered Oct4 or control siRNAs. Anti-GATA6 antibodies (red), anti-SSEA-4 antibodies (green), genomic DNA (blue). ( f, g ) Analysis of IFN-α and TNF-α induction in human PBMCs at 4 or 24 h post-treatment with mock (PBS), β-gal siRNA plus PTD-DRBD or plus Lipofection, as indicated. 10 μ/ml Imiquimod Imiquimod or 10 μg/ml LPS was used as a positive control for IFN-α or TNF-α, respectively. ( h ) Nasal and tracheal expressing ROSA26R-Luciferase transgenic mice were live animal imaged on day 0. Randomized groups of luciferase expressing mice were then treated with PBS, PTD-DRBD plus Luc siRNA or control GFP (Con) siRNA and monitored daily for luciferase expression, as indicated. Scale is in photons/s/cm 2 /sr. ( i ) Graph of percent Luciferase knockdown mice from (h) above. Luciferase expression was normalized to mock each day, error bar indicates s.e.m., n = 3 for each group with two luciferase readings performed per mouse per day.

    Journal: Nature biotechnology

    Article Title: Efficient siRNA Delivery into Primary Cells by Peptide Transduction-dsRNA Binding Domain (PTD-DRBD) Fusion Protein

    doi: 10.1038/nbt.1541

    Figure Lengend Snippet: PTD-DRBD mediated RNAi responses ( a ) Fluorescent microscopy analysis of H9 hESCs constitutively expressing GFP treated with with PTD-DRBD delivered GFP2 siRNA at 2 days post-addition. Black line outlines hESC colony on mouse feeder cell background. ( b ) Oct4 immunoblot analysis in HUES9 hESCs treated with mock (PBS), PTD-DRBD delivered Oct4 or control siRNAs at 2 days post-addition. ( c ) Cell division curve of human HUES9 embryonic stem cells treated with mock (PBS), PTD-DRBD delivered Oct4 or control siRNAs, as indicated. ( d ) Immunohistochemistry analysis of Oct4 and SSEA4 expression in HUES9 hESCs at 2 days post-treatment with mock (PBS), PTD-DRB delivered Oct4 or control siRNAs. Anti-Oct4 antibodies (red), anti-SSEA-4 antibodies (green), genomic DNA (blue). ( e ) Immuno-histochemistry analysis of GATA6 and SSEA4 expression in HUES9 hESCs at 10 days post-treatment with mock (PBS), PTD-DRB delivered Oct4 or control siRNAs. Anti-GATA6 antibodies (red), anti-SSEA-4 antibodies (green), genomic DNA (blue). ( f, g ) Analysis of IFN-α and TNF-α induction in human PBMCs at 4 or 24 h post-treatment with mock (PBS), β-gal siRNA plus PTD-DRBD or plus Lipofection, as indicated. 10 μ/ml Imiquimod Imiquimod or 10 μg/ml LPS was used as a positive control for IFN-α or TNF-α, respectively. ( h ) Nasal and tracheal expressing ROSA26R-Luciferase transgenic mice were live animal imaged on day 0. Randomized groups of luciferase expressing mice were then treated with PBS, PTD-DRBD plus Luc siRNA or control GFP (Con) siRNA and monitored daily for luciferase expression, as indicated. Scale is in photons/s/cm 2 /sr. ( i ) Graph of percent Luciferase knockdown mice from (h) above. Luciferase expression was normalized to mock each day, error bar indicates s.e.m., n = 3 for each group with two luciferase readings performed per mouse per day.

    Article Snippet: PTD-DRBD expression utilized BL21 codon plus (DH3) E.coli (Strategene) cells were transformed with pPTD-DRBD, cultured at 37°C in LB, then at 25°C for 12 h after induction with 400 μM IPTG.

    Techniques: Microscopy, Expressing, Immunohistochemistry, Positive Control, Luciferase, Transgenic Assay, Mouse Assay

    Competitive binding of cannabinoid ligands on BL21-107 membranes expressing CB2. The assay was performed in triplicate as described in Materials and Methods.

    Journal:

    Article Title: Expression of human peripheral cannabinoid receptor for structural studies

    doi: 10.1110/ps.051550305

    Figure Lengend Snippet: Competitive binding of cannabinoid ligands on BL21-107 membranes expressing CB2. The assay was performed in triplicate as described in Materials and Methods.

    Article Snippet: E. coli protease-deficient expression strain BL21 (DE3) was purchased from Stratagene.

    Techniques: Binding Assay, Expressing

    Ligand-binding assay on E. coli BL21-107 membranes

    Journal:

    Article Title: Expression of human peripheral cannabinoid receptor for structural studies

    doi: 10.1110/ps.051550305

    Figure Lengend Snippet: Ligand-binding assay on E. coli BL21-107 membranes

    Article Snippet: E. coli protease-deficient expression strain BL21 (DE3) was purchased from Stratagene.

    Techniques: Ligand Binding Assay

    Time course of CB2-107 expression in E. coli . ( A ) An overnight culture of BL21-107 cells was used to inoculate several incubation flasks containing equal volumes of media. Induction was started when cell density reached OD 600 = 0.4. Cell samples were

    Journal:

    Article Title: Expression of human peripheral cannabinoid receptor for structural studies

    doi: 10.1110/ps.051550305

    Figure Lengend Snippet: Time course of CB2-107 expression in E. coli . ( A ) An overnight culture of BL21-107 cells was used to inoculate several incubation flasks containing equal volumes of media. Induction was started when cell density reached OD 600 = 0.4. Cell samples were

    Article Snippet: E. coli protease-deficient expression strain BL21 (DE3) was purchased from Stratagene.

    Techniques: Expressing, Incubation

    Saturation binding of [ 3 H] CP-55,940 on BL21-107 membranes expressing CB2. The assay was performed in triplicate as described in Materials and Methods.

    Journal:

    Article Title: Expression of human peripheral cannabinoid receptor for structural studies

    doi: 10.1110/ps.051550305

    Figure Lengend Snippet: Saturation binding of [ 3 H] CP-55,940 on BL21-107 membranes expressing CB2. The assay was performed in triplicate as described in Materials and Methods.

    Article Snippet: E. coli protease-deficient expression strain BL21 (DE3) was purchased from Stratagene.

    Techniques: Binding Assay, Expressing

    SDS-PAGE analysis of purified protein recombinant SllB in E. coli BL21 cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.

    Journal: International Journal of Molecular Medicine

    Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

    doi: 10.3892/ijmm.2012.890

    Figure Lengend Snippet: SDS-PAGE analysis of purified protein recombinant SllB in E. coli BL21 cells. Lane M: Takara Protein Marker; lane 1, SDS-PAGE analysis of the recombinant S-layer protein before purification; lane 2, SDS-PAGE analysis of the purified recombinant protein.

    Article Snippet: 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

    Techniques: SDS Page, Purification, Recombinant, Marker

    A transmission electron microscopic observation on the E. coli BL21 with recombinant protein. (A) Normal E. coli BL21 was treated as control. (B) The E. coli BL21 cells recombinant S-layer protein. (C) Crystal lattice structures on surface of the E. coli BL21 cells recombinant S-layer protein.

    Journal: International Journal of Molecular Medicine

    Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

    doi: 10.3892/ijmm.2012.890

    Figure Lengend Snippet: A transmission electron microscopic observation on the E. coli BL21 with recombinant protein. (A) Normal E. coli BL21 was treated as control. (B) The E. coli BL21 cells recombinant S-layer protein. (C) Crystal lattice structures on surface of the E. coli BL21 cells recombinant S-layer protein.

    Article Snippet: 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

    Techniques: Transmission Assay, Recombinant

    Expression of sllB in E. coli BL21 cells. (A) SDS-PAGE analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; lane M represent Takara Protein Marker (Broad). (B) Western blot analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; M, Takara Protein Marker (Broad). Lane M1, Precision plus protein standards; lane M2, perfect protein marker. Note the band pointed with arrows is the recombinant S-layer protein.

    Journal: International Journal of Molecular Medicine

    Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification

    doi: 10.3892/ijmm.2012.890

    Figure Lengend Snippet: Expression of sllB in E. coli BL21 cells. (A) SDS-PAGE analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; lane M represent Takara Protein Marker (Broad). (B) Western blot analysis of rSllB protein; lane 1, the whole cell lysate of E. coli BL21 cells containing pET28a(+); lane 2, the supernatant of cells containing pET28a(+); lane 3, the pellet of cells containing pET28a(+); lane 4, the whole cell lysate of E. coli BL21 cells containing pET28a(+)- sllB ; lane 5, the supernatant of cell containing pET28a(+)- sllB ; lane 6, the pellet of cells containing pET28a(+)- sllB ; M, Takara Protein Marker (Broad). Lane M1, Precision plus protein standards; lane M2, perfect protein marker. Note the band pointed with arrows is the recombinant S-layer protein.

    Article Snippet: 2.0 (Takara, no. DV801A), from which 0.5 μl was used to transform 100 μl of E. coli BL21 (DE3, Stratagene).

    Techniques: Expressing, SDS Page, Marker, Western Blot, Recombinant