Journal: Diagnostics

Article Title: Innovative DendrisChips ® Technology for a Syndromic Approach of In Vitro Diagnosis: Application to the Respiratory Infectious Diseases

doi: 10.3390/diagnostics8040077

Figure Lengend Snippet: Pathogen bacterial strains used to evaluate the specificity of the designed probes from hypervariable rDNA regions in their genomes.

Article Snippet: Mycoplasma pneumoniae , ATCC 29342D , ATCC *.

Techniques:

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Schematic representation of M. pneumoniae M129 P1 gene and its four gene fragments; P1-I, P1-II, P1-III and P1-IV. Each bar represents the position of UGA codons that codes for tryptophan. To express these fragments, UGA codons were modified to UGG. Fragments were amplified using a set of forward (F) and reverse primers (R).

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Modification, Amplification

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: SDS-PAGE and Western blot analysis of recombinant M. pneumoniae P1 proteins fragments. (A) Coomassie blue stained SDS-PAGE analysis of rP1-I, rP1-II, rP1-III and rP1-IV in E. coli extract. The fragments were expressed in pET28b vector and protein production was induced with IPTG in E. coli. (B) Western blot analysis of induced and uninduced P1 protein fragments rP1-I, rP1-II, rP1-IV (i) and rP1-III (ii), showing reactivity with anti-6X His antibody. (C) Coomassie blue stained SDS-PAGE analysis of Ni 2+ -NTA purified P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV. (D) Western blot analysis of purified P1 protein fragments rP1-I, rP1-II, rP1-III and rP1-IV showing reactivity with anti-6X His antibody. Lane Marker: Molecular mass marker (kDa); Arrows indicate position of expressed protein.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: SDS Page, Western Blot, Recombinant, Staining, Plasmid Preparation, Purification, Marker

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Western blot and ELISA analysis of M. pneumoniae lysate and Cross reactivity of Pab (rP1-I) and Pab (rP1-IV). Reactivity of P1 (170 kDa) with anti-P1 protein fragments antibody Pab (rP1-I), Pab (rP1-II), Pab (rP1-III) & Pab (rP1-IV) rose in Rabbit by western blotting (A) and by ELISA (B) . (C) & (D) Immuno blot analysis of rP1-I, rP1-II, rP1-III and rP1-IV fragments with Pab (rP1-I) and Pab (rP1-IV) showing their cross reactivity with respective sera. Lane Marker: Molecular mass marker (kDa).

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Marker

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Recombinant P1 protein fragments are recognized by anti- M. pneumoniae antibody and by sera of M. pneumoniae infected patients. (A) (I) Coomassie blue stained SDS-PAGE analysis of purified M. pneumoniae P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV. Immuno blot analysis of purified P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV using anti- M. pneumoniae antibody (II) and using pooled sera of M. pneumoniae infected patients (III). (B) Immuno blot analysis of purified M. pneumoniae P1 protein fragments rP1-I, rP1-II, rP1-III and rP1-IV with several sera of M. pneumoniae infected patients. PM: Prestained protein marker; PC: positive control; NC: Negative control; Numbers over the blot indicate serial number of sera of M. pneumoniae infected patients tested for these experiments.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Recombinant, Infection, Staining, SDS Page, Purification, Marker, Positive Control, Negative Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Comparative ELISA analysis of recombinant P1 protein fragments with sera of M. pneumoniae infected patients. Reactivity of purified M. pneumoniae P1 proteins fragments with 25 sera of M. pneumoniae infected patients by ELISA (A) , with 16 healthy patient sera (B) and average values of both A & B (C) . Number on top of column indicates serial number of sera of M. pneumoniae infected patients tested for these experiments.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Infection, Purification

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: IFM adhesion assay of M. pneumoniae (A-E). The M. pneumoniae attached to the HEp-2 cells were detected by either anti- M. pneumoniae antibody or antibodies rose in rabbits. The detecting antibodies were added after fixation with methanol. (A) anti- M. pneumoniae antibody (positive control), (B) Pab (rP1-I), (C) Pab (rP1-II), (D) Pab (rP1-III), (E) Pab (rP1-IV). IFM surface exposure assay of M. pneumoniae (F-J) . In this assay the detecting antibodies were added before the methanol fixation. (F) anti- M. pneumoniae antibody (positive control), (G) Pab (rP1-I), (H) Pab (rP1-II), (I) Pab (rP1-III), (J) Pab (rP1-IV). Negative controls: (K) mycoplasmas alone (Without Pabs), (L) Pabs alone (Without mycoplasmas). Bar, 2 μm.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Cell Adhesion Assay, Positive Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: IFM adhesion inhibition assay. M. pneumoniae were pre-incubated with either anti- M. pneumoniae antibodies or antibodies rose in rabbits in different dilutions (1:50, 1:100, 1:200, 1:500) before infection of the HEp-2 cells. These antibodies were: (A-D) anti- M. pneumoniae antibody (positive control), (E-H) Pab (rP1-I), (I-L) Pab (rP1-IV), (M) Pab (rP1-II) (N) Pab (rP1-III) (O) Without antibody, (P) pre-immune serum. Bar, 2 μm.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Inhibition, Incubation, Infection, Positive Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Primer sequence used to amplify all four fragments of M. pneumoniae M129 P1 gene

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: (A) Cartoon representation of the lowest energy structure on the van der Waals surface. (B) Sequence of HuPrP(90–231, M129, Q212P) protein. The elements of secondary structure are shown. (C) Structural details of α 3 helix in the family of 20 lowest energy structures from Met205 to Arg220 in the Q212P mutant (left, pdb id 2KUN) and WT HuPrP C (right, pdb id 1QM1) . Residues Pro212 and Gln212 are presented in pink. The r.m.s.d. for backbone atoms in residues between Met205 and Arg220 in both ensembles is 0.7 Å.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques: Sequencing, Mutagenesis

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: NMR restrains and structural statistics for the ensemble of 20 lowest energy structures of HuPrP(90-231, M129, Q212P) protein.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques:

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: (A) Type of NOE used in structure calculations of HuPrP(90–231, M129, Q212P) protein. (B) Enlarged region between Tyr218 and Tyr225. Glu221 and Ser222 do not exhibit any long-range NOE contacts. (C) Schematic presentation of long-range NOE contacts (|i-j|>5) between residues in helices a 3 and a 4 . For clarity, only inter-helical NOE contacts are shown.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques:

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: (A) HuPrP(90–231, M129, Q212P) (pdb id 2KUN, this work). (B) WT HuPrP(90–231, M129) (pdb id 1QM1) . (C) Elk PrP (pdb id 1XYW) . (D) Bank vole PrP (pdb id 2K56) . (E) Tammar wallaby PrP (pdb id 2KFL) .

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques:

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: (A) Carton presentation of α 2 , α 3 and α 4 helices with mutual orientation of Phe175 and Gln217 in the Q212P mutant. (B) Carton presentation of α 2 and α 3 helices with mutual orientation of Phe175 and Gln217 in the WT protein. (C) The mutual orientation of α 2 and α 3 helices with indicated inter-helical angle in Q212P mutant. (D) The mutual orientation of α 2 and α 3 helices with indicated inter-helical angle in WT protein. (E) Structural organization of β 2 -α 2 loop and α 3 and α 4 helices in Q212P mutant. (F) Structural organization of β 2 -α 2 loop and α 3 helix in WT protein.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques: Mutagenesis

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: Inter-helical angles and distances between helices α 2 and α 3 in high-resolution structures of PrP C proteins a .

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques:

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: Distances between Tyr225 and residues within β 2 –α 2 loop region and α 3 helix.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques:

Journal: PLoS ONE

Article Title: NMR Structure of the Human Prion Protein with the Pathological Q212P Mutation Reveals Unique Structural Features

doi: 10.1371/journal.pone.0011715

Figure Lengend Snippet: (A) Structural details of β 2 α 2 α 3 α 4 region (161–228) of 20 lowest energy structures for HuPrP(90–231, M129, Q212P) mutant. (B) Structural details of β 2 α 2 α 3 region (161–228) of 20 lowest energy structures for HuPrP(90–231, M129, R220K) (pdb id 1E1U) mutant . (C) Structural details of β 2 α 2 α 3 region (161–228) of 20 lowest energy structures for HuPrP(90–231, M129, E200K) (pdb id 1FO7) mutant . In all three panels the point mutation is indicated in magenta (left). Top view of the hydrophobic core composed of aromatic amino acid residues (center). Hydrophobicity surface is presented on the right, where red color indicates hydrophobic and blue color represents hydrophilic surface.

Article Snippet: All NMR experiments used for structure calculation were performed on 13 C, 15 N double labeled HuPrP(90-231, M129, Q212P) protein on Varian VNMRS 800 MHz NMR spectrometer using triple 1 H/ 13 C/ 15 N resonance cold probe-head with inverse detection at 298 K. NMR sample consisted in a 0.8 mM concentration of 13 C, 15 N-labeled protein in sodium acetate buffer.

Techniques: Mutagenesis

Journal: iScience

Article Title: A tetracationic porphyrin with dual anti-prion activity

doi: 10.1016/j.isci.2023.107480

Figure Lengend Snippet:

Article Snippet: pET-41 huPrP90-231 (M129) , Provided by B. Caughey, Rocky Mountain Laboratories, Hamilton, MT, USA , N/A.

Techniques: Western Blot, Virus, Recombinant, Saline, Reverse Transcription, Bicinchoninic Acid Protein Assay, Generated, Software, Imaging