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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 "Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation"

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation

Journal: Molecular bioSystems

doi: 10.1039/c2mb25051a

Abl kinase domain and Abl SH3-SH2-Kinase single chain multi-domain expression in bacteria and their purification analysed by SDS-PAGE and Western blot. (A) SDS-PAGE of Abl kinase: whole-cell lysate (lane 1); soluble proteins after centrifuge (lane 2); proteins bound on Co 2+ resin (lane3); retaining proteins on Co 2+ resin after thrombin cleavage (lane 4). (B) Tyrosine phosphorylation status: whole-cell lysate of BL21 with GroEL expression (lane 1); whole-cell lysate of BL21 with GroEL and Abl kinase domain co-expression (lane 2); proteins bound on Co 2+ resin (lane 3); proteins on Co 2+ resin after thrombin cleavage (lane 4); proteins eluted from the affinity resin (lane 5); protein from Lane 5 after CIP treatment (lane 6); dephosphorylated Abl kinase domain after purification (lane 7). The upper panel is Coomassie blue stained SDS-PAGE, the lower panel is an immunoblot with anti-phosphotyrosine antibody. (C) SDS-PAGE of Abl SH3-SH2-kinase: cell lysate before and after centrifuge (land 1 and 2); proteins remained on and eluted from the TALON resin after thrombin cleavage (land 3 and 4); after ionic exchange purification (lane 5); after both ionic exchange and size exclusion purification (lane 6); NMR sample before and after measurement (lane 7 and 8).
Figure Legend Snippet: Abl kinase domain and Abl SH3-SH2-Kinase single chain multi-domain expression in bacteria and their purification analysed by SDS-PAGE and Western blot. (A) SDS-PAGE of Abl kinase: whole-cell lysate (lane 1); soluble proteins after centrifuge (lane 2); proteins bound on Co 2+ resin (lane3); retaining proteins on Co 2+ resin after thrombin cleavage (lane 4). (B) Tyrosine phosphorylation status: whole-cell lysate of BL21 with GroEL expression (lane 1); whole-cell lysate of BL21 with GroEL and Abl kinase domain co-expression (lane 2); proteins bound on Co 2+ resin (lane 3); proteins on Co 2+ resin after thrombin cleavage (lane 4); proteins eluted from the affinity resin (lane 5); protein from Lane 5 after CIP treatment (lane 6); dephosphorylated Abl kinase domain after purification (lane 7). The upper panel is Coomassie blue stained SDS-PAGE, the lower panel is an immunoblot with anti-phosphotyrosine antibody. (C) SDS-PAGE of Abl SH3-SH2-kinase: cell lysate before and after centrifuge (land 1 and 2); proteins remained on and eluted from the TALON resin after thrombin cleavage (land 3 and 4); after ionic exchange purification (lane 5); after both ionic exchange and size exclusion purification (lane 6); NMR sample before and after measurement (lane 7 and 8).

Techniques Used: Expressing, Purification, SDS Page, Western Blot, Staining, Nuclear Magnetic Resonance

3) 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

4) Product Images from "Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen"

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen

Journal: American Journal of Translational Research

doi:

Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of
Figure Legend Snippet: Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of

Techniques Used: Expressing, Purification, Recombinant, SDS Page, Plasmid Preparation, Marker

5) Product Images from "Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae"

Article Title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Journal: Brazilian Journal of Medical and Biological Research

doi: 10.1590/S0100-879X2012007500077

Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.
Figure Legend Snippet: Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.

Techniques Used: Expressing, Transformation Assay, Plasmid Preparation, SDS Page, Marker, Western Blot

6) Product Images from "Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen"

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen

Journal: American Journal of Translational Research

doi:

Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of
Figure Legend Snippet: Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of

Techniques Used: Expressing, Purification, Recombinant, SDS Page, Plasmid Preparation, Marker

7) 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

8) Product Images from "Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli"

Article Title: Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli

Journal: Journal of Bacteriology

doi:

Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained
Figure Legend Snippet: Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained

Techniques Used: Expressing, SDS Page, Polyacrylamide Gel Electrophoresis, Staining

9) 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

10) Product Images from "Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli"

Article Title: Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli

Journal: Journal of Bacteriology

doi:

Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained
Figure Legend Snippet: Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained

Techniques Used: Expressing, SDS Page, Polyacrylamide Gel Electrophoresis, Staining

11) 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

12) 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

13) 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

14) 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

15) Product Images from "Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation"

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation

Journal: Molecular bioSystems

doi: 10.1039/c2mb25051a

Abl kinase domain and Abl SH3-SH2-Kinase single chain multi-domain expression in bacteria and their purification analysed by SDS-PAGE and Western blot. (A) SDS-PAGE of Abl kinase: whole-cell lysate (lane 1); soluble proteins after centrifuge (lane 2); proteins bound on Co 2+ resin (lane3); retaining proteins on Co 2+ resin after thrombin cleavage (lane 4). (B) Tyrosine phosphorylation status: whole-cell lysate of BL21 with GroEL expression (lane 1); whole-cell lysate of BL21 with GroEL and Abl kinase domain co-expression (lane 2); proteins bound on Co 2+ resin (lane 3); proteins on Co 2+ resin after thrombin cleavage (lane 4); proteins eluted from the affinity resin (lane 5); protein from Lane 5 after CIP treatment (lane 6); dephosphorylated Abl kinase domain after purification (lane 7). The upper panel is Coomassie blue stained SDS-PAGE, the lower panel is an immunoblot with anti-phosphotyrosine antibody. (C) SDS-PAGE of Abl SH3-SH2-kinase: cell lysate before and after centrifuge (land 1 and 2); proteins remained on and eluted from the TALON resin after thrombin cleavage (land 3 and 4); after ionic exchange purification (lane 5); after both ionic exchange and size exclusion purification (lane 6); NMR sample before and after measurement (lane 7 and 8).
Figure Legend Snippet: Abl kinase domain and Abl SH3-SH2-Kinase single chain multi-domain expression in bacteria and their purification analysed by SDS-PAGE and Western blot. (A) SDS-PAGE of Abl kinase: whole-cell lysate (lane 1); soluble proteins after centrifuge (lane 2); proteins bound on Co 2+ resin (lane3); retaining proteins on Co 2+ resin after thrombin cleavage (lane 4). (B) Tyrosine phosphorylation status: whole-cell lysate of BL21 with GroEL expression (lane 1); whole-cell lysate of BL21 with GroEL and Abl kinase domain co-expression (lane 2); proteins bound on Co 2+ resin (lane 3); proteins on Co 2+ resin after thrombin cleavage (lane 4); proteins eluted from the affinity resin (lane 5); protein from Lane 5 after CIP treatment (lane 6); dephosphorylated Abl kinase domain after purification (lane 7). The upper panel is Coomassie blue stained SDS-PAGE, the lower panel is an immunoblot with anti-phosphotyrosine antibody. (C) SDS-PAGE of Abl SH3-SH2-kinase: cell lysate before and after centrifuge (land 1 and 2); proteins remained on and eluted from the TALON resin after thrombin cleavage (land 3 and 4); after ionic exchange purification (lane 5); after both ionic exchange and size exclusion purification (lane 6); NMR sample before and after measurement (lane 7 and 8).

Techniques Used: Expressing, Purification, SDS Page, Western Blot, Staining, Nuclear Magnetic Resonance

16) Product Images from "Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli"

Article Title: Tetrachloroethene Dehalogenase from Dehalospirillum multivorans: Cloning, Sequencing of the Encoding Genes, and Expression of the pceA Gene in Escherichia coli

Journal: Journal of Bacteriology

doi:

Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained
Figure Legend Snippet: Expression of pceA from D. multivorans in E. coli BL21 (DE3) as analyzed by SDS-PAGE. Strains of E. coli BL21 (DE3) harboring different plasmids were induced by IPTG. Cell extracts were analyzed by SDS–12% PAGE and subsequently stained

Techniques Used: Expressing, SDS Page, Polyacrylamide Gel Electrophoresis, Staining

17) Product Images from "Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae"

Article Title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Journal: Brazilian Journal of Medical and Biological Research

doi: 10.1590/S0100-879X2012007500077

Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.
Figure Legend Snippet: Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.

Techniques Used: Expressing, Transformation Assay, Plasmid Preparation, SDS Page, Marker, Western Blot

18) Product Images from "Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen"

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen

Journal: American Journal of Translational Research

doi:

Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of
Figure Legend Snippet: Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of

Techniques Used: Expressing, Purification, Recombinant, SDS Page, Plasmid Preparation, Marker

19) 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

20) Product Images from "Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae"

Article Title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Journal: Brazilian Journal of Medical and Biological Research

doi: 10.1590/S0100-879X2012007500077

Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.
Figure Legend Snippet: Expression of rDer f 5 in Escherichia coli BL21 cells. E. coli BL21 cells were transformed with either pET28a(+)-Der f 5 or empty vector pET28a(+) as control. A , SDS-PAGE analysis of the rDer f 5 protein. Lane M 1 = TaKaRa protein marker (Broad); lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = supernatant of cells containing pET28a; lane 3 = pellet of cells containing pET28a; lane 4 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5; lane 5 = supernatant of cells containing pET28a(+)-Der f 5; lane 6 = pellet of cells containing pET28a(+)-Der f 5. B , Western blotting analysis of the rDer f 5 protein. Lane M 2 = Precision Plus Protein Standards; lane 1 = whole cell lysate of E. coli BL21 cells containing pET28a; lane 2 = whole cell lysate of E. coli BL21 cells containing pET28a(+)-Der f 5. Arrows point to the band of rDer f 5.

Techniques Used: Expressing, Transformation Assay, Plasmid Preparation, SDS Page, Marker, Western Blot

21) Product Images from "Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen"

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen

Journal: American Journal of Translational Research

doi:

Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of
Figure Legend Snippet: Expression and purification of recombinant protein Der f 21. A. SDS-PAGE analysis of the protein expressed from the pCold-TF- Der f 21 recombinant plasmid in E. coli BL21 cells. Lane M , TaKaRa Protein Marker (Broad); Lane 1 , the whole cell lysate of

Techniques Used: Expressing, Purification, Recombinant, SDS Page, Plasmid Preparation, Marker

Related Articles

Ligation:

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation
Article Snippet: .. Plasmid containing SH3-SH2-kinase, pMAL/SH32(C2A)kin and plasmid containing kinase domain for ligation, pMAL/SspDnaAblCkinase were simply transformed into E. coli BL21 for recombinant protein expression. .. For pTBX1/AblSH32G_(C2A), BL21-codonplus(DE3)RIPL (Stratagene) were used.

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).

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).

Expressing:

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: .. E. coli BL21 was transformed with plasmid pET28a (+)-sllB and protein expression was induced with IPTG. .. A single band from SDS-PAGE ( ) and western blotting ( ) was observed on whole cell lysate, soluble protein fraction, and insoluble protein fraction, confirming the predicted molecular weight for the S-layer protein.

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation
Article Snippet: .. Plasmid containing SH3-SH2-kinase, pMAL/SH32(C2A)kin and plasmid containing kinase domain for ligation, pMAL/SspDnaAblCkinase were simply transformed into E. coli BL21 for recombinant protein expression. .. For pTBX1/AblSH32G_(C2A), BL21-codonplus(DE3)RIPL (Stratagene) were used.

Sequencing:

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen
Article Snippet: .. After sequencing, the verified pCold-TF-Der f 21 plasmids were transformed into E. coli BL21, which were then grown on LB plates. .. Protein expression was induced by IPTG treatment.

Transformation Assay:

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: .. E. coli BL21 was transformed with plasmid pET28a (+)-sllB and protein expression was induced with IPTG. .. A single band from SDS-PAGE ( ) and western blotting ( ) was observed on whole cell lysate, soluble protein fraction, and insoluble protein fraction, confirming the predicted molecular weight for the S-layer protein.

Article Title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae
Article Snippet: .. E. coli BL21 was then transformed with plasmid pET28a-(+)-Der f 5 and the protein expressed was induced with IPTG at 37°C for 3 h. Absorbance at 600 nm was measured for 0.562 and 1.259 before and after induction. ..

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen
Article Snippet: .. Transformed E. coli BL21 were induced to express the recombinant protein by treatment with IPTG. ..

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen
Article Snippet: .. After sequencing, the verified pCold-TF-Der f 21 plasmids were transformed into E. coli BL21, which were then grown on LB plates. .. Protein expression was induced by IPTG treatment.

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation
Article Snippet: .. Plasmid containing SH3-SH2-kinase, pMAL/SH32(C2A)kin and plasmid containing kinase domain for ligation, pMAL/SspDnaAblCkinase were simply transformed into E. coli BL21 for recombinant protein expression. .. For pTBX1/AblSH32G_(C2A), BL21-codonplus(DE3)RIPL (Stratagene) were used.

Recombinant:

Article Title: Cloning, expression, and analysis of a cDNA coding for the Dermatophagoides farinae group 21 (Der f 21) allergen
Article Snippet: .. Transformed E. coli BL21 were induced to express the recombinant protein by treatment with IPTG. ..

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation
Article Snippet: .. Plasmid containing SH3-SH2-kinase, pMAL/SH32(C2A)kin and plasmid containing kinase domain for ligation, pMAL/SspDnaAblCkinase were simply transformed into E. coli BL21 for recombinant protein expression. .. For pTBX1/AblSH32G_(C2A), BL21-codonplus(DE3)RIPL (Stratagene) were used.

Plasmid Preparation:

Article Title: Cloning of the surface layer gene sllB from Bacillus sphaericus ATCC 14577 and its heterologous expression and purification
Article Snippet: .. E. coli BL21 was transformed with plasmid pET28a (+)-sllB and protein expression was induced with IPTG. .. A single band from SDS-PAGE ( ) and western blotting ( ) was observed on whole cell lysate, soluble protein fraction, and insoluble protein fraction, confirming the predicted molecular weight for the S-layer protein.

Article Title: Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae
Article Snippet: .. E. coli BL21 was then transformed with plasmid pET28a-(+)-Der f 5 and the protein expressed was induced with IPTG at 37°C for 3 h. Absorbance at 600 nm was measured for 0.562 and 1.259 before and after induction. ..

Article Title: Abl Kinase Constructs Expressed in Bacteria: facilitation of structural and functional studies including segmental labeling by expressed protein ligation
Article Snippet: .. Plasmid containing SH3-SH2-kinase, pMAL/SH32(C2A)kin and plasmid containing kinase domain for ligation, pMAL/SspDnaAblCkinase were simply transformed into E. coli BL21 for recombinant protein expression. .. For pTBX1/AblSH32G_(C2A), BL21-codonplus(DE3)RIPL (Stratagene) were used.

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  • 85
    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: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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: 93/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene e coli bl21 gold
    In vivo interactions of MBP-UreD with (UreABC) 3 , UreB, (UreAC) 3 , and UreBΔ1-19. (A) SDS-PAGE depicting the interactions of MBP-UreD with (UreABC) 3 , UreB, and (UreAC) 3 . E. coli <t>BL21-Gold(DE3)</t> cells were co-transformed with pCDF-MBP-UreD (encoding
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    Stratagene e coli bl21 de3 ripl cells
    A biochemical approach to dissect portal function. (a) The plasmids containing g20 variants (WT or mutants) were transformed into E. coli <t>BL21</t> (DE3) <t>RIPL</t> strain for IPTG induced expression of the respective gp20 protein (green). (b) The E. coli cells
    E Coli Bl21 De3 Ripl Cells, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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

    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

    In vivo interactions of MBP-UreD with (UreABC) 3 , UreB, (UreAC) 3 , and UreBΔ1-19. (A) SDS-PAGE depicting the interactions of MBP-UreD with (UreABC) 3 , UreB, and (UreAC) 3 . E. coli BL21-Gold(DE3) cells were co-transformed with pCDF-MBP-UreD (encoding

    Journal: Biochemistry

    Article Title: The Function of UreB in Klebsiella aerogenes Urease

    doi: 10.1021/bi2011064

    Figure Lengend Snippet: In vivo interactions of MBP-UreD with (UreABC) 3 , UreB, (UreAC) 3 , and UreBΔ1-19. (A) SDS-PAGE depicting the interactions of MBP-UreD with (UreABC) 3 , UreB, and (UreAC) 3 . E. coli BL21-Gold(DE3) cells were co-transformed with pCDF-MBP-UreD (encoding

    Article Snippet: UreBΔ1-19 was purified from E. coli BL21-Gold(DE3) harboring pUreBΔ1-19 by a process similar to that for UreB.

    Techniques: In Vivo, SDS Page, Transformation Assay

    A biochemical approach to dissect portal function. (a) The plasmids containing g20 variants (WT or mutants) were transformed into E. coli BL21 (DE3) RIPL strain for IPTG induced expression of the respective gp20 protein (green). (b) The E. coli cells

    Journal: Journal of molecular biology

    Article Title: Structure-Function Analysis of the DNA Translocating Portal of the Bacteriophage T4 Packaging Machine

    doi: 10.1016/j.jmb.2013.10.011

    Figure Lengend Snippet: A biochemical approach to dissect portal function. (a) The plasmids containing g20 variants (WT or mutants) were transformed into E. coli BL21 (DE3) RIPL strain for IPTG induced expression of the respective gp20 protein (green). (b) The E. coli cells

    Article Snippet: E. coli BL21 (DE3) RIPL cells containing the gp20 recombinant plasmids was induced with isopropyl-β-D-thio-galactoside (IPTG; 1 mM) for 20 min at 37°C.

    Techniques: Transformation Assay, Expressing