scholarly journals Introduction of an Aliphatic Ketone into Recombinant Proteins in a Bacterial Strain that Overexpresses an Editing-Impaired Leucyl-tRNA Synthetase

ChemBioChem ◽  
2009 ◽  
Vol 10 (13) ◽  
pp. 2188-2190 ◽  
Author(s):  
Yi Tang ◽  
Pin Wang ◽  
James A. Van Deventer ◽  
A. James Link ◽  
David A. Tirrell
Keyword(s):  
Recombinant Proteins ◽  
Bacterial Strain ◽  
Trna Synthetase ◽  
Aliphatic Ketone

scholarly journals Detecting the nature and solving the crystal structure of a contaminant protein from an opportunistic pathogen

2020 ◽  
Vol 76 (9) ◽  
pp. 392-397 ◽  
Author(s):  
Riccardo Pederzoli ◽  
Delia Tarantino ◽  
Louise J. Gourlay ◽  
Antonio Chaves-Sanjuan ◽  
Martino Bolognesi
Keyword(s):  
Crystal Structure ◽  
Escherichia Coli ◽  
Structure Analysis ◽  
Recombinant Proteins ◽  
Bacterial Strain ◽  
Burkholderia Pseudomallei ◽  
Opportunistic Pathogen ◽  
Laboratory Environment ◽  
E Coli ◽  
Different Sources

The unintentional crystallization of contaminant proteins in the place of target recombinant proteins is sporadically reported, despite the availability of stringent expression/purification protocols and of software for the detection of contaminants. Typically, the contaminant protein originates from the expression organism (for example Escherichia coli), but in rare circumstances contaminants from different sources have been reported. Here, a case of contamination from a Serratia bacterial strain that occurred while attempting to crystallize an unrelated protein from Burkholderia pseudomallei (overexpressed in E. coli) is presented. The contamination led to the unintended crystallization and structure analysis of a cyanate hydratase from a bacterial strain of the Serratia genus, an opportunistic enterobacterium that grows under conditions similar to those of E. coli and that is found in a variety of habitats, including the laboratory environment. In this context, the procedures that were adopted to identify the contaminant based on crystallographic data only are presented and the crystal structure of Serrata spp. cyanate hydratase is briefly discussed.

scholarly journals Staphylococcus aureus Protection-Related Type 3 Cell-Mediated Immune Response Elicited by Recombinant Proteins and GM-CSF DNA Vaccine

2021 ◽  
Author(s):  
Kamila R. Santos ◽  
Fernando N. Souza ◽  
Eduardo R. Sanchez ◽  
Camila F. Batista ◽  
Luiza C. Reis ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Immune Response ◽  
Dna Vaccine ◽  
Recombinant Proteins ◽  
Trna Synthetase ◽  
Gm Csf ◽  
Interleukin 17A ◽  
Cell Mediated Immune Response ◽  
Macrophage Colony ◽  
Type 3

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A+ cells among overall CD44+ (memory), T CD4+, CD4+ T CD44+ CD27-, γδ TCR, γδ TCR+ CD44+ CD27+ and TCRVγ4+ cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulates TH2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4+ and CD4+ TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus.

scholarly journals Staphylococcus aureus Protection-Related Type 3 Cell-Mediated Immune Response Elicited by Recombinant Proteins and GM-CSF DNA Vaccine

Vaccines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 899
Author(s):  
Kamila R. Santos ◽  
Fernando N. Souza ◽  
Eduardo M. Ramos-Sanchez ◽  
Camila F. Batista ◽  
Luiza C. Reis ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Immune Response ◽  
Dna Vaccine ◽  
Recombinant Proteins ◽  
Interferon Γ ◽  
Trna Synthetase ◽  
Gm Csf ◽  
Interleukin 17A ◽  
Macrophage Colony ◽  
Type 3

Staphylococcus aureus mastitis remains a major challenge for dairy farming. Here, 24 mice were immunized and divided into four groups: G1: control; G2: Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) DNA vaccine; G3: F0F1 ATP synthase subunit α (SAS), succinyl-diaminopimelate (SDD), and cysteinyl-tRNA synthetase (CTS) recombinant proteins; and G4: SAS+SDD+CTS plus GM-CSF DNA vaccine. The lymphocyte subpopulations, and the intracellular interleukin-17A (IL-17A) and interferon-γ production in the draining lymph node cells were immunophenotyped by flow cytometry. The immunophenotyping and lymphocyte proliferation was determined in spleen cells cultured with and without S. aureus stimulus. Immunization with S. aureus recombinant proteins generated memory cells in draining lymph nodes. Immunization with the three recombinant proteins plus GM-CSF DNA led to an increase in the percentage of IL-17A+ cells among overall CD44+ (memory), T CD4+, CD4+ T CD44+ CD27−, γδ TCR, γδ TCR+ CD44+ CD27+, and TCRVγ4+ cells. Vaccination with S. aureus recombinant proteins associated with GM-CSF DNA vaccine downregulated TH2 immunity. Immunization with the three recombinant proteins plus the GM-CSF DNA led to a proliferation of overall memory T, CD4+, and CD4+ TEM cells upon S. aureus stimulus. This approach fostered type 3 immunity, suggesting the development of a protective immune response against S. aureus.

Sign in / Sign up

Export Citation Format

Share Document