e. coli strain dh-5α  (Promega)

Journal: Veterinary Sciences

Article Title: Immunogenicity Characterization of the Recombinant gI Protein Fragment from Pseudorabies Virus and an Evaluation of Its Diagnostic Use in Pigs

doi: 10.3390/vetsci10080506

Figure Lengend Snippet: Expression and identification of truncated gI protein. ( A ) SDS-PAGE gel results of target proteins expressed in E. coli after 0.3 and 1.0 mM IPTG induction for 7 h at 37 °C. pET-30(a): inducible expression band of empty vector; pET-30(a)-gI: the induced expression of an approximately 38 kDa band of recombinant expression plasmid. ( B ) Western blot analysis was conducted with pig standard positive serum against PRV as the primary antibody; pET-30(a): inducible expression band of empty vector; pET-30(a)-gI: pig standard positive serum against PRV as the primary antibody. ( C ) predicted secondary structure of the monomeric form of the truncated gI peptides generated using Protean software.

Article Snippet: The pBluescript and pET-30a (+) vectors, Escherichia coli ( E. coli ) strains DH 5α and BL 21 (DE3), Luria–Bertani (LB) medium, X-Gal, and isopropyl β-d-thiogalactoside (IPTG) used in this study were purchased from Tiangen, Beijing, China.

Techniques: Expressing, SDS Page, Plasmid Preparation, Recombinant, Western Blot, Generated, Software

Journal: bioRxiv

Article Title: The Synechocystis sp. PCC 6803 open reading frame slr0201 that is homologous to sdhC from Archaea codes for a [2Fe-2S] protein

doi: 10.1101/2021.09.23.461530

Figure Lengend Snippet: Heterologous overexpression of slr0201 carrying a 9 x His-tag in E. coli strain BL21 (DE3) plysS under the T7 RNA polymerase promoter. Overexpression of slr0201 was induced by addition of 1 mM IPTG at time 0. A : Protein profile of the whole cell lysate extracted at the times indicated after IPTG addition. The overexpressed Slr0201 has been indicated by an arrow. M: molecular size marker. B : Staining intensity of the Slr0201 region as a function of time after IPTG addition.

Article Snippet: The E. coli strains DH 5α and BL21 (DE3) plys S (Stratagene) were used for plasmid propagation and overexpression of the recombinant protein, respectively.

Techniques: Over Expression, Marker, Staining

Journal: bioRxiv

Article Title: The Synechocystis sp. PCC 6803 open reading frame slr0201 that is homologous to sdhC from Archaea codes for a [2Fe-2S] protein

doi: 10.1101/2021.09.23.461530

Figure Lengend Snippet: Distribution of Slr0201upon overexpression of slr0201 in E. coli at 25°C and 37°C. The SDS-PAGE gel stained with Coomassie Brilliant Blue shows polypeptide profiles of the whole cell lysate , the supernatant , the inclusion body pellet , and the membrane fraction . Slr0201 in the inclusion body fraction (lane 3 in each panel) has been indicated by arrows. Molecular weights of the markers (M) have been indicated.

Article Snippet: The E. coli strains DH 5α and BL21 (DE3) plys S (Stratagene) were used for plasmid propagation and overexpression of the recombinant protein, respectively.

Techniques: Over Expression, SDS Page, Staining

Journal: bioRxiv

Article Title: The Synechocystis sp. PCC 6803 open reading frame slr0201 that is homologous to sdhC from Archaea codes for a [2Fe-2S] protein

doi: 10.1101/2021.09.23.461530

Figure Lengend Snippet: Analysis of expression of slr0201 in Synechocystis sp. PCC 6803. Soluble ( , , ) and membrane proteins ( , , ) from the wild type ( , ), the Δ slr0201 mutant ( , ), and the slr0201 + -His strain ( , ) were separated by SDS-PAGE gel ( A ). After transfer to nitrocellulose membrane, the expressed slr0201 products were immunoreacted with either antibodies raised against Slr0201 ( B) or a commercial His-tag monoclonal antibody for expression of the inserted, His-tagged slr0201 ( C ). The sample loaded in lane “M” in panel B and C is the purified Slr0201 overproduced in E. coli strain BL21 (DE3) plysS carrying the plasmid pET16S0201. Panel D shows the purified, solubilized Slr0201 inclusion bodies used as antigen to raise Slr0201 antibodies (lane 1) as well as the Slr0201 antibodies before (lane 2) and after (lane 3) affinity purification.

Article Snippet: The E. coli strains DH 5α and BL21 (DE3) plys S (Stratagene) were used for plasmid propagation and overexpression of the recombinant protein, respectively.

Techniques: Expressing, Mutagenesis, SDS Page, Purification, Plasmid Preparation, Affinity Purification

Journal: Biomolecules

Article Title: Galectin Domain Containing Protein from Haemonchus contortus Modulates the Immune Functions of Goat PBMCs and Regulates CD4+ T-Helper Cells In Vitro

doi: 10.3390/biom10010116

Figure Lengend Snippet: Expression and purification of rHc-GDC. Lane M: Standard molecular weight protein marker; Lane 1: Recombinant expression vector before induction of Isopropyl-β-d- thiogalactopyranoside (IPTG); Lane 2–5: Expression after IPTG induction at different time points; Lane 6: Pre-purified rHc-GDC; Lane 7: Purified rHc-GDC.

Article Snippet: Transformation of recombinant plasmid (pMD19-T/GDC) into E. coli DH 5α strain (Invitrogen) was performed and cultured in ampicillin containing Luria Bertini (LB) medium.

Techniques: Expressing, Purification, Molecular Weight, Marker, Recombinant, Plasmid Preparation

Journal: Biomolecules

Article Title: Galectin Domain Containing Protein from Haemonchus contortus Modulates the Immune Functions of Goat PBMCs and Regulates CD4+ T-Helper Cells In Vitro

doi: 10.3390/biom10010116

Figure Lengend Snippet: Expression of rHc-GDC protein in adult H. contortus by immunofluorescence assay. ( A , B ) nuclei stained with DAPI (blue); ( C , D ) target protein localization in adult worms using Cy3-conjugated secondary antibody; ( E , F ) merge image of DAPI and Cy3. No florescence was observed in control. Scale bar 40 μm.

Article Snippet: Transformation of recombinant plasmid (pMD19-T/GDC) into E. coli DH 5α strain (Invitrogen) was performed and cultured in ampicillin containing Luria Bertini (LB) medium.

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: Biomolecules

Article Title: Galectin Domain Containing Protein from Haemonchus contortus Modulates the Immune Functions of Goat PBMCs and Regulates CD4+ T-Helper Cells In Vitro

doi: 10.3390/biom10010116

Figure Lengend Snippet: Effects of rHc-GDC on PBMC migration. Cells were treated with PBS (control), purified pET32a protein, and serial concentrations of rHc-GDC. Then, the random migration was determined. Statistical differences between standard error mean of three independent experiments ( n = 5) were calculated using one-way ANOVA and marked ** when p < 0.01, *** when p < 0.001 and ns when no significant difference was found.

Article Snippet: Transformation of recombinant plasmid (pMD19-T/GDC) into E. coli DH 5α strain (Invitrogen) was performed and cultured in ampicillin containing Luria Bertini (LB) medium.

Techniques: Migration, Control, Purification

Journal: Biomolecules

Article Title: Galectin Domain Containing Protein from Haemonchus contortus Modulates the Immune Functions of Goat PBMCs and Regulates CD4+ T-Helper Cells In Vitro

doi: 10.3390/biom10010116

Figure Lengend Snippet: Influence of rHc-GDC on nitric oxide production by PBMCs in vitro. Cells were incubated with PBS, pET32a, and serial concentrations of rHc-GDC protein for 24 h in 37 °C and 5% CO 2 . The nitrate concentration in the PBMCs was measured by using the Griess assay. Statistical differences between standard error mean of three independent experiments were calculated using one way ANOVA and marked * when p < 0.05, ** when p < 0.01, *** when p < 0.001, and ns when no significant difference was found.

Article Snippet: Transformation of recombinant plasmid (pMD19-T/GDC) into E. coli DH 5α strain (Invitrogen) was performed and cultured in ampicillin containing Luria Bertini (LB) medium.

Techniques: In Vitro, Incubation, Concentration Assay, Griess Assay