Structured Review

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: 91/100, based on 92 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Wild-type Escherichia coli grows on the chitin disaccharide, N,N?-diacetylchitobiose, by expressing the cel operon"

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

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

doi:

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.
Figure Legend 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.

Techniques Used: Mutagenesis

2) Product Images from "A New Amidohydrolase from Bordetella or Alcaligenes Strain FB188 with Similarities to Histone Deacetylases"

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

Journal: Journal of Bacteriology

doi: 10.1128/JB.186.8.2328-2339.2004

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.
Figure Legend 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.

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

3) Product Images from "Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage"

Article Title: Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage

Journal: PLoS ONE

doi: 10.1371/journal.pone.0014702

pVII tag-modified helper phages are structurally and functionally identical to normal helper phages, and donate a defined phenotype to pVII. ( A ) Schematic illustration of the pV, pVII, pIX genomic junctions in the M13 genome framed by the unique BsrG I/ SnaB I RE sites. The three tag modifications, their site of insertion and physical characteristics are given. Isoelectric point (pI) as well as average charge (§) was computed using ProtParam ( http://ca.expasy.org/ ). ( B ) AviTag-pVII functionality as BirA substrate assessed by virion binding to magnetic SA beads and detection with an anti-M13 Ab (BirA enzyme activity provided by E. coli XL1-Blue or AVB100FmkII). ( C ) FLAG-pVII functionality as assessed by virion binding to immobilized anti-FLAG M2 and M5 mAbs and detection with anti-M13 Ab. ( D ) HIS 6 -pVII functionality assessed by virion binding to magnetic IMAC beads and detection with an anti-M13 Ab. Virion binding to the beads was done without ( gray bars ) or with ( black bar ) 300 mM imidazole in the binding buffer. ( E ) HIS 6 -pVII functionality assessed as in D, employing HIS 6 -pVII tagged pVIII phagemid display (pGALD8mFN, unpublished ) virions ( left part ). Specific IMAC bead capture efficiency assessed by titration of imidazole-eluted pVIII phagemid virions after bead capture, shown as percent of in-put (% recovery = out-put (cfu ampR )/in-put (cfu ampR )×100), right part .
Figure Legend Snippet: pVII tag-modified helper phages are structurally and functionally identical to normal helper phages, and donate a defined phenotype to pVII. ( A ) Schematic illustration of the pV, pVII, pIX genomic junctions in the M13 genome framed by the unique BsrG I/ SnaB I RE sites. The three tag modifications, their site of insertion and physical characteristics are given. Isoelectric point (pI) as well as average charge (§) was computed using ProtParam ( http://ca.expasy.org/ ). ( B ) AviTag-pVII functionality as BirA substrate assessed by virion binding to magnetic SA beads and detection with an anti-M13 Ab (BirA enzyme activity provided by E. coli XL1-Blue or AVB100FmkII). ( C ) FLAG-pVII functionality as assessed by virion binding to immobilized anti-FLAG M2 and M5 mAbs and detection with anti-M13 Ab. ( D ) HIS 6 -pVII functionality assessed by virion binding to magnetic IMAC beads and detection with an anti-M13 Ab. Virion binding to the beads was done without ( gray bars ) or with ( black bar ) 300 mM imidazole in the binding buffer. ( E ) HIS 6 -pVII functionality assessed as in D, employing HIS 6 -pVII tagged pVIII phagemid display (pGALD8mFN, unpublished ) virions ( left part ). Specific IMAC bead capture efficiency assessed by titration of imidazole-eluted pVIII phagemid virions after bead capture, shown as percent of in-put (% recovery = out-put (cfu ampR )/in-put (cfu ampR )×100), right part .

Techniques Used: Modification, Binding Assay, Activity Assay, Titration

4) Product Images from "Expression of Active Recombinant Human Tissue-Type Plasminogen Activator by Using In Vivo Polyhydroxybutyrate Granule Display ▿"

Article Title: Expression of Active Recombinant Human Tissue-Type Plasminogen Activator by Using In Vivo Polyhydroxybutyrate Granule Display ▿

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.01543-10

SDS-PAGE analysis of the PhaP-rPA fusion protein. Lane M, protein marker; lane 1, control; lane 2, lysate of E. coli XL1-Blue (pHBS01/pSPAr); lane 3, PHB granules isolated from E. coli XL1-Blue (pHBS01/pSPAr).
Figure Legend Snippet: SDS-PAGE analysis of the PhaP-rPA fusion protein. Lane M, protein marker; lane 1, control; lane 2, lysate of E. coli XL1-Blue (pHBS01/pSPAr); lane 3, PHB granules isolated from E. coli XL1-Blue (pHBS01/pSPAr).

Techniques Used: SDS Page, Recombinase Polymerase Amplification, Marker, Isolation

Cell growth and PHB accumulation of recombinant E. coli . The strains were cultivated in LB medium supplemented with 0.5% glucose. (A) Comparison of PHB content and cell mass in E. coli BL21/pHBS01 and XL1-Blue/pHBS01. (B) Fluorescence micrograph
Figure Legend Snippet: Cell growth and PHB accumulation of recombinant E. coli . The strains were cultivated in LB medium supplemented with 0.5% glucose. (A) Comparison of PHB content and cell mass in E. coli BL21/pHBS01 and XL1-Blue/pHBS01. (B) Fluorescence micrograph

Techniques Used: Recombinant, Fluorescence

Western blot analysis of the PhaP-rPA fusion protein. Lane M, protein marker; lane 1, control; lane 2, PHB granules isolated from E. coli XL1-Blue (pHBS01/pSPAr).
Figure Legend Snippet: Western blot analysis of the PhaP-rPA fusion protein. Lane M, protein marker; lane 1, control; lane 2, PHB granules isolated from E. coli XL1-Blue (pHBS01/pSPAr).

Techniques Used: Western Blot, Recombinase Polymerase Amplification, Marker, Isolation

Fibrin plate analysis of rPA activity. Equal amounts of the samples were spotted on the fibrin plate. (A) Cell debris from E. coli XL1-Blue (pSCP/pSPAr). (B) Isolated PHB granules from E. coli XL1-Blue (pHBS01/pSPAr).
Figure Legend Snippet: Fibrin plate analysis of rPA activity. Equal amounts of the samples were spotted on the fibrin plate. (A) Cell debris from E. coli XL1-Blue (pSCP/pSPAr). (B) Isolated PHB granules from E. coli XL1-Blue (pHBS01/pSPAr).

Techniques Used: Recombinase Polymerase Amplification, Activity Assay, Isolation

Related Articles

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: Antigenic Importance of the Carboxy-Terminal Beta-Strand of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein
Article Snippet: .. E. coli strains XL1-Blue (Stratagene, La Jolla, Calif.) and DH5α were used as hosts for generating N gene mutations after PCR mutagenesis and for general-purpose cloning, respectively. ..

Article Title: Molecular Dissection of Phage Endolysin
Article Snippet: .. E. coli strain XL1-Blue (Stratagene) was used for all cloning procedures, and BL21(DE3) strain (Lucigen) of this bacterium was used in all experiments involving protein expression. .. E. coli strains were grown in Luria-Bertani medium (HiMedia) having 100 μg/ml ampicillin at 37 °C and with constant shaking at 200 rpm unless specified otherwise.

Centrifugation:

Article Title: Antigenic Importance of the Carboxy-Terminal Beta-Strand of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein
Article Snippet: Cellular debris was removed by centrifugation at 1,500 × g for 10 min, and the clarified supernatant was used as crude virus stock. .. E. coli strains XL1-Blue (Stratagene, La Jolla, Calif.) and DH5α were used as hosts for generating N gene mutations after PCR mutagenesis and for general-purpose cloning, respectively.

Amplification:

Article Title: Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose ▿
Article Snippet: .. Plasmids were amplified in E. coli strain XL1-Blue (Stratagene, La Jolla, CA). .. E. coli was grown on Luria-Bertani plates or in liquid Terrific broth medium for the isolation of plasmids ( ).

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Paragraph title: PCR amplification and construction of expression constructs ... All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Construct:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies. .. An R. prowazekii cosmid clone bank constructed in E. coli DH1 was used in transport screening experiments ( , ).

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Paragraph title: PCR amplification and construction of expression constructs ... All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Infection:

Article Title: Antigenic Importance of the Carboxy-Terminal Beta-Strand of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein
Article Snippet: To prepare the PRRSV isolate PA8 , MARC-145 cells were infected with an MOI of 5 PFU/cell. .. E. coli strains XL1-Blue (Stratagene, La Jolla, Calif.) and DH5α were used as hosts for generating N gene mutations after PCR mutagenesis and for general-purpose cloning, respectively.

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: For uptake assays, rickettsial suspensions were concentrated so that rickettsiae derived from 8 g of infected yolk sac were present in 1 ml. .. E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies.

Expressing:

Article Title: Molecular Dissection of Phage Endolysin
Article Snippet: .. E. coli strain XL1-Blue (Stratagene) was used for all cloning procedures, and BL21(DE3) strain (Lucigen) of this bacterium was used in all experiments involving protein expression. .. E. coli strains were grown in Luria-Bertani medium (HiMedia) having 100 μg/ml ampicillin at 37 °C and with constant shaking at 200 rpm unless specified otherwise.

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: Paragraph title: Protein Expression & Purification ... The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

Article Title: Interaction of the Sliding Clamp ?-Subunit and Hda, a DnaA-Related Protein
Article Snippet: E. coli strains XL1-Blue (Stratagene) and TOP10 (Invitrogen) were used as hosts in site-directed mutagenesis and subcloning of PCR inserts. .. BL21(DE3)pLyS (Novagen) and TB1 (New England Biolabs) were used as hosts for protein expression.

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Paragraph title: PCR amplification and construction of expression constructs ... All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Transformation Assay:

Article Title: Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose ▿
Article Snippet: S. cerevisiae cells were plated onto MYD after transformation with plasmids. .. Plasmids were amplified in E. coli strain XL1-Blue (Stratagene, La Jolla, CA).

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. The sequence of the purified plasmid was confirmed by automated DNA sequencing (Sequencing Facility, University of California, Berkeley).

Derivative Assay:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: For uptake assays, rickettsial suspensions were concentrated so that rickettsiae derived from 8 g of infected yolk sac were present in 1 ml. .. E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies.

Article Title: Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose ▿
Article Snippet: The S. cerevisiae strains used in this work were derived from the xylose-fermenting strain RWB217 ( ) and are listed in Table . .. Plasmids were amplified in E. coli strain XL1-Blue (Stratagene, La Jolla, CA).

Transfection:

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. Cell stocks for each mutant were then expanded and DNA purified using an endotoxin-free Qiagen Highspeed Midiprep or Maxiprep Kit prior to transfection into S2 cells.

Cell Culture:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies. .. E. coli strains were cultured in Luria-Bertani (LB) medium at 37°C.

Article Title: Production of recombinant antibody fragments in Bacillus megaterium
Article Snippet: Production of scFvs in E. coli The D1.3 scFv was produced in shaking flasks according to Dübel et al. [ ] using the vector pOPE101 [ ] and the E. coli strain XL1-Blue MRF' (Stratagene, Amsterdam, Netherland). .. Briefly, 300 mL 2 × TY + 100 mM glucose + 100 μg/mL ampicillin were inoculated with an overnight culture yield to O.D.600 nm = 0.1 and cultured at 37°C and 250 rpm.

Generated:

Article Title: Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L.) Taub)
Article Snippet: Two cDNA libraries were generated: an "early" seed library (15, 20, and 25 DAF, library I), and a "late" seed library (30, 35, and 40 DAF, library II). .. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene, Los Angeles, CA).

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: Tyr216 mutations were generated by PCR using the pBipTBM plasmid and primers encoding 10–17 bases upstream and downstream of the mutation. .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

DNA Sequencing:

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

Polymerase Chain Reaction:

Article Title: Antigenic Importance of the Carboxy-Terminal Beta-Strand of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein
Article Snippet: .. E. coli strains XL1-Blue (Stratagene, La Jolla, Calif.) and DH5α were used as hosts for generating N gene mutations after PCR mutagenesis and for general-purpose cloning, respectively. ..

Article Title: Escherichia coli Isolate for Studying Colonization of the Mouse Intestine and Its Application to Two-Component Signaling Knockouts
Article Snippet: .. The tetR tetA cassette was isolated by PCR from the transposon Tn 10 in the E. coli strain XL1-Blue (Stratagene). pML8 is pAS07 with mcherry (taken from pRSETb-mCherry [ ]) in place of gfpmut3.1 . .. The plasmids pAS07 and pML8 were integrated into the MP1 genome at the phage lambda attachment site and verified to be present in single copy by PCR as described in reference , resulting in MP13 and MP7, respectively.

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: Tyr216 mutations were generated by PCR using the pBipTBM plasmid and primers encoding 10–17 bases upstream and downstream of the mutation. .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Paragraph title: PCR amplification and construction of expression constructs ... All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Injection:

Article Title: Transcriptional profiling reveals multifunctional roles for transferrin in the honeybee, Apis mellifera
Article Snippet: Septic injury One microliter of either bacterial or yeast cells (10 × concentrated overnight cultures in sterile bee Ringer) were injected into the thorax of ice-chilled 3–4-day old bees using a 25 µl Hamilton syringe attached to a dispenser. .. We used E. coli strain XL1-Blue from Stratagen ( www.stratagene.com ) and S. cerevisiae strain Dip2 ( ).

Recombinant:

Article Title: Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L.) Taub)
Article Snippet: .. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene, Los Angeles, CA). .. Excised plasmids were plated using SOLR cells (Stratagene, Los Angeles, CA).

Article Title: Studies on the nonmevalonate pathway to terpenes: The role of the GcpE (IspG) protein
Article Snippet: .. The resulting plasmid pBSxylBCDF was electrotransformed into E. coli strain XL1-Blue, yielding the recombinant strain XL1-pBSxylBispCDF. .. The E. coli ispE gene (GenBank accession no. AE000219 ) was amplified from bp position 5720 to 6571 by PCR with the use of chromosomal E. coli DNA as template and the oligonucleotides ychB Xho I and ychB Kpn I as primers (Table ).

In Vivo:

Article Title: Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L.) Taub)
Article Snippet: .. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene, Los Angeles, CA). .. Excised plasmids were plated using SOLR cells (Stratagene, Los Angeles, CA).

Mutagenesis:

Article Title: Antigenic Importance of the Carboxy-Terminal Beta-Strand of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein
Article Snippet: .. E. coli strains XL1-Blue (Stratagene, La Jolla, Calif.) and DH5α were used as hosts for generating N gene mutations after PCR mutagenesis and for general-purpose cloning, respectively. ..

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: Tyr216 mutations were generated by PCR using the pBipTBM plasmid and primers encoding 10–17 bases upstream and downstream of the mutation. .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

Isolation:

Article Title: Escherichia coli Isolate for Studying Colonization of the Mouse Intestine and Its Application to Two-Component Signaling Knockouts
Article Snippet: .. The tetR tetA cassette was isolated by PCR from the transposon Tn 10 in the E. coli strain XL1-Blue (Stratagene). pML8 is pAS07 with mcherry (taken from pRSETb-mCherry [ ]) in place of gfpmut3.1 . .. The plasmids pAS07 and pML8 were integrated into the MP1 genome at the phage lambda attachment site and verified to be present in single copy by PCR as described in reference , resulting in MP13 and MP7, respectively.

Article Title: Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L.) Taub)
Article Snippet: Poly A+ RNA was isolated using an Oligotex mRNA Mini Kit (Qiagen, Los Angeles, CA). cDNA was prepared from polyA+ enriched, pooled samples of equivalent amounts of total RNA from each time point. .. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene, Los Angeles, CA).

Article Title: Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose ▿
Article Snippet: Plasmids were amplified in E. coli strain XL1-Blue (Stratagene, La Jolla, CA). .. E. coli was grown on Luria-Bertani plates or in liquid Terrific broth medium for the isolation of plasmids ( ).

Subcloning:

Article Title: Interaction of the Sliding Clamp ?-Subunit and Hda, a DnaA-Related Protein
Article Snippet: .. E. coli strains XL1-Blue (Stratagene) and TOP10 (Invitrogen) were used as hosts in site-directed mutagenesis and subcloning of PCR inserts. .. BL21(DE3)pLyS (Novagen) and TB1 (New England Biolabs) were used as hosts for protein expression.

Size-exclusion Chromatography:

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. Protein secreted by S2 cells was purified using anion exchange (DEAE), His-tag affinity (TALON), and size exclusion chromatography (GE) as described previously for WT TβM.

Purification:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: Rickettsiae were purified from the yolk sacs of embryonated hen eggs as described previously ( ) and were suspended in a sucrose-phosphate-glutamate-magnesium solution (SPG-Mg; 0.218 M sucrose, 3.76 mM KH2 PO4 , 7.1 mM K2 HPO4 , 4.9 mM potassium glutamate, and 10 mM MgCl2 ). .. E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies.

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. The sequence of the purified plasmid was confirmed by automated DNA sequencing (Sequencing Facility, University of California, Berkeley).

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Purified PCR products corresponding to NS3, NS5A and NS5A-T were restricted using the restriction enzymes Bam HI/Hind III, Nde I/Hind III and Nde I/Hind III, respectively and ligated into pET11a vector that was restricted using same restriction enzymes. .. All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Sequencing:

Article Title: Escherichia coli Isolate for Studying Colonization of the Mouse Intestine and Its Application to Two-Component Signaling Knockouts
Article Snippet: The plasmid pAS07 (A. Siryaporn and M. Goulian, unpublished data) consists of the integration vector pCAH63 ( ) with the uidA gene replaced with a sequence consisting of tetR and an operon fusion of tetA with gfpmut3.1 (Clontech). .. The tetR tetA cassette was isolated by PCR from the transposon Tn 10 in the E. coli strain XL1-Blue (Stratagene). pML8 is pAS07 with mcherry (taken from pRSETb-mCherry [ ]) in place of gfpmut3.1 .

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit.

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: The identity of plasmids was confirmed by nucleotide sequencing. .. All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

IA:

Article Title: Molecular Analysis of Group B Protective Surface Protein, a New Cell Surface Protective Antigen of Group B Streptococci
Article Snippet: Wild GBS strains were H4A-0126 (Ia/R1, BPS), H4A-0148 (Ia,/R1, BPS) and B176 (Ia, BPS). .. The Escherichia coli strains XL1-Blue MRF and XLOLR were obtained from a commercial source (Stratagene).

Plasmid Preparation:

Article Title: Escherichia coli Isolate for Studying Colonization of the Mouse Intestine and Its Application to Two-Component Signaling Knockouts
Article Snippet: The plasmid pAS07 (A. Siryaporn and M. Goulian, unpublished data) consists of the integration vector pCAH63 ( ) with the uidA gene replaced with a sequence consisting of tetR and an operon fusion of tetA with gfpmut3.1 (Clontech). .. The tetR tetA cassette was isolated by PCR from the transposon Tn 10 in the E. coli strain XL1-Blue (Stratagene). pML8 is pAS07 with mcherry (taken from pRSETb-mCherry [ ]) in place of gfpmut3.1 .

Article Title: Production of recombinant antibody fragments in Bacillus megaterium
Article Snippet: .. Production of scFvs in E. coli The D1.3 scFv was produced in shaking flasks according to Dübel et al. [ ] using the vector pOPE101 [ ] and the E. coli strain XL1-Blue MRF' (Stratagene, Amsterdam, Netherland). .. Briefly, 300 mL 2 × TY + 100 mM glucose + 100 μg/mL ampicillin were inoculated with an overnight culture yield to O.D.600 nm = 0.1 and cultured at 37°C and 250 rpm.

Article Title: Analysis of cDNA libraries from developing seeds of guar (Cyamopsis tetragonoloba (L.) Taub)
Article Snippet: .. Phagemids containing cDNA inserts were in vivo excised from the recombinant Uni-ZAP XR vector using ExAssist helper phage and the E. coli strain XL1-Blue MRF' (Stratagene, Los Angeles, CA). .. Excised plasmids were plated using SOLR cells (Stratagene, Los Angeles, CA).

Article Title: The Interdomain Long-Range Electron Transfer Becomes Rate-Limiting in the Y216A Variant of Tyramine ?-Monooxygenase
Article Snippet: .. The reverse primers were complementary: Y216A: G GAG ACC ACG GCC TGG TGT CAC G Y216I: CC AGT CAG GAG ACC ACG ATC TGG TGT CAC GTT CAG C Y216W: C AGT CAG GAG ACC ACG TGG TGG TGT CAC GTT CAG C The resulting altered pBipTBM plasmid was transformed into Escherichia coli strain XL1 Blue (Stratagene) cells and purified using a Qiagen Highspeed Miniprep kit. .. The sequence of the purified plasmid was confirmed by automated DNA sequencing (Sequencing Facility, University of California, Berkeley).

Article Title: Interaction of the Sliding Clamp ?-Subunit and Hda, a DnaA-Related Protein
Article Snippet: E. coli strains XL1-Blue (Stratagene) and TOP10 (Invitrogen) were used as hosts in site-directed mutagenesis and subcloning of PCR inserts. .. Expression vector pET16b was from Novagen, and expression vector pMAL was from New England Biolabs.

Article Title: Studies on the nonmevalonate pathway to terpenes: The role of the GcpE (IspG) protein
Article Snippet: .. The resulting plasmid pBSxylBCDF was electrotransformed into E. coli strain XL1-Blue, yielding the recombinant strain XL1-pBSxylBispCDF. .. The E. coli ispE gene (GenBank accession no. AE000219 ) was amplified from bp position 5720 to 6571 by PCR with the use of chromosomal E. coli DNA as template and the oligonucleotides ychB Xho I and ychB Kpn I as primers (Table ).

Article Title: Over-expression and characterization of NS3 and NS5A of Hepatitis C virus genotype 3a
Article Snippet: Purified PCR products corresponding to NS3, NS5A and NS5A-T were restricted using the restriction enzymes Bam HI/Hind III, Nde I/Hind III and Nde I/Hind III, respectively and ligated into pET11a vector that was restricted using same restriction enzymes. .. All genetic manipulation was performed in Escherichia coli strain XL1-Blue (Stratagene, USA).

Article Title: Expanding the Versatility of Phage Display I: Efficient Display of Peptide-Tags on Protein VII of the Filamentous Phage
Article Snippet: Plasmids, bacterial strains, phage and materials The phOx-BSA and pSEX81 phagemid vector harboring a phOx-BSA specific affinity matured human scFv was kindly provided by Affitech Research AS (Oslo, Norway). .. The E. coli strains XL1-Blue (recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F proAB lacI q ZΔM15 Tn10 (Tetr )] and CJ236 (FΔ(HindIII)::cat (Tra+Pil+Camr )/ung-1 relA1 dut-1 thi-1 spoT1 mcrA ) were purchased from Stratagene (LaJolla, CA, USA) and New England Biolabs (Ipswich, MA, USA), respectively.

Selection:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies. .. Where appropriate for selection of E. coli transformants, ampicillin was added to a final concentration of 50 μg/ml.

Produced:

Article Title: Production of recombinant antibody fragments in Bacillus megaterium
Article Snippet: .. Production of scFvs in E. coli The D1.3 scFv was produced in shaking flasks according to Dübel et al. [ ] using the vector pOPE101 [ ] and the E. coli strain XL1-Blue MRF' (Stratagene, Amsterdam, Netherland). .. Briefly, 300 mL 2 × TY + 100 mM glucose + 100 μg/mL ampicillin were inoculated with an overnight culture yield to O.D.600 nm = 0.1 and cultured at 37°C and 250 rpm.

Concentration Assay:

Article Title: S-Adenosylmethionine Transport in Rickettsia prowazekii
Article Snippet: E. coli strain XL1-Blue (Stratagene, La Jolla, Calif.) was used as the standard recipient in these studies. .. Where appropriate for selection of E. coli transformants, ampicillin was added to a final concentration of 50 μg/ml.

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  • 91
    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: 91/100, 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 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    e coli strain xl1 blue mr - by Bioz Stars, 2020-02
    91/100 stars
      Buy from Supplier

    99
    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: 99/100, 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 99 stars, based on 43 article reviews
    Price from $9.99 to $1999.99
    e coli xl1 blue - by Bioz Stars, 2020-02
    99/100 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