Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Vector map of recombinant pCR™8/GW/TOPO™ (Gateway, Invitrogen) with cloned com1 (pCR8- com1 ).

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Plasmid Preparation, Recombinant, Clone Assay

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Cloned com1 . Amplification of com1 (762 bp) without the 5′-terminal signal sequence (1–60 bp) but including the 3′-terminal stop codon (759–762) from genomic DNA of C. burnetii Nine Mile Phase II RSA 439. Arrows show com1 CBU_1910 F and com1 CBU_1910 R primers used for amplification.

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Clone Assay, Amplification, Sequencing

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Vector map of recombinant expression vector pET300/NT-DEST (Invitrogen) with cloned com1 cloned into (pET300- com1 ).

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Plasmid Preparation, Recombinant, Expressing, Clone Assay

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Cloned and expressed Com1 protein. ( a ) Insertion of com1 into pET300/NT-DEST vector for expression of recombinant Com1 of 29.9 kDa with His-tag. ( b ) Expression of His-tagged com1 as recombinant Com1 protein. ATG, vector encoded start codon; RBS, vector encoded ribosome binding site; 6xHis, vector encoded His-tag. Axis counting base pairs in relation to com1 .

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Clone Assay, Plasmid Preparation, Expressing, Recombinant, Binding Assay

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Analysis of purified recombinant Com1 protein with His-tag. Ni-NTA purified recombinant Com1 protein (predicted molecular weight 29.9 kDa, 30 µg per lane) was separated using a 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). ( a ) For Western blot analysis mouse anti-6xHis monoclonal antibody (1:5000, Clontech Laboratories, Inc., Mountain View, CA, USA) was used and traced with alkaline phosphatase labeled goat anti-mouse secondary antibody (1:5000, Sigma-Aldrich, St. Louis, MO, USA). ( c ) Recombinant Com1 protein was visualized using colloidal Coomassie Blue. Lane ( a , c ), purified recombinant Com1 protein; Lane ( b ), protein marker with size in kDa (Thermo Fisher Scientific, Waltham, MA, USA).

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Purification, Recombinant, Molecular Weight, Polyacrylamide Gel Electrophoresis, SDS Page, Western Blot, Labeling, Marker

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Q fever status of sera from sheep, goats and cattle collected during Q fever outbreaks, from apparently healthy herds or supplied by Friedrich–Loeffler Institute, Federal Research Institute for Animal Health (FLI) National Reference Laboratories (NRL).

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Enzyme-linked Immunosorbent Assay, Bacteria, Recombinant

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Statistical values for the recombinant Com1-ELISA at the determined threshold value of 0.32 for sheep sera. The contingency table shows the number of true and false negative as well as true and false positive results for sheep serum samples.

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Statistical values for the recombinant Com1-ELISA at the determined threshold value of 0.22 for caprine sera. The contingency table shows the number of true and false negative as well as true and false positive results for goat serum samples.

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Microorganisms

Article Title: Evaluation of the Diagnostic Potential of Recombinant Coxiella burnetii Com1 in an ELISA for the Diagnosis of Q Fever in Sheep, Goats and Cattle

doi: 10.3390/microorganisms8081235

Figure Lengend Snippet: Statistical values for the recombinant Com1-ELISA at the determined threshold value of 0.18 for cattle sera. The contingency table shows the number of true and false negative as well as true and false positive results for cattle serum samples.

Article Snippet: His-tagged recombinant Com1 protein was detected using anti-6xHis monoclonal mouse antibody (1:5000 in BlueBlock PF, Clontech) and traced with an alkaline phosphatase (AP) conjugated goat anti-mouse monoclonal antibody (1:5000 in BlueBlock PF, Sigma Aldrich, St. Louis, MO, USA) for 1 h at room temperature and shaken.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: BIOCHEMICAL ANALYSES OF TRANSCRIPTIONAL REGULATORY MECHANISMS IN A CHROMATIN CONTEXT

doi: 10.1016/j.ymeth.2006.11.004

Figure Lengend Snippet: (A) Relaxed plasmid DNA was assembled into chromatin without (−) or with (+) histone H1. Core histones were titrated onto the DNA template to form nucleosomes. The core histone to DNA (CH/DNA) mass ratios are indicated. The resulting change in DNA topology was resolved using 1%TBE agarose gel electrophoresis. Nucleic acids were visualized with SYBR gold staining and STORM imaging. The positions of nicked, relaxed and supercoiled (SC) DNA are illustrated. (B) Chromatin was assembled in the absence (−) or presence (+) of histone H1 and digested with micrococcal nuclease as described previously. The nucleosome repeat lengths have been calculated for each assembly and are as follows: no H1 - 170 (+/− 9 bp), dH1 - 186 (+/− 8 bp), and CEM H1 – 189 (+/− 10 bp). Base pair markers from Fermentas (100 bp DNA ladder plus) are indicated to the right.

Article Snippet: Supercoiled plasmid DNA should be separated from nicked DNA using agarose gel preparative-cell electrophoresis (Bio-Rad).

Techniques: Plasmid Preparation, Agarose Gel Electrophoresis, Staining, Imaging