dna sequence coding for the c3 domain of adhesin p1  (GenScript corporation)

Journal: Vaccines

Article Title: Three-Component Subunit Vaccine Induces Protective Immunity Against Mycoplasma pneumoniae in Mice

doi: 10.3390/vaccines14040330

Figure Lengend Snippet: Recombinant protein design, construction, and purity validation. ( A ) Schematic diagram of recombinant protein constructs. Mutation sites are indicated by short black lines. ( B – H ) SDS-PAGE, Western blot, and Superdex™ 200 Increase 10/300 GL purification results for recombinant proteins. ( B ) P1 (~165 kDa); ( C ) P40/90 complex (~117 kDa); ( D ) CARDS-WT (~69 kDa); ( E ) CARDS-E132A (~69 kDa); ( F ) CARDS-E132Q (~69 kDa); ( G ) CARDS-H36A (~69 kDa); ( H ) CARDS-R10A (~69 kDa). Left lane: protein marker; right lane: purified recombinant protein.

Article Snippet: The coding sequences for P1 (GenBank I D: X06871 ), P40 (GenBank ID: AF125204 ), P90 (GenBank ID: AF125205 ), and CARDS-WT (GenBank ID: AY189944 ) were retrieved from the NCBI database based on the Mycoplasma pneumoniae standard strain ATCC M129.

Techniques: Recombinant, Biomarker Discovery, Construct, Mutagenesis, SDS Page, Western Blot, Purification, Marker

Journal: Vaccines

Article Title: Three-Component Subunit Vaccine Induces Protective Immunity Against Mycoplasma pneumoniae in Mice

doi: 10.3390/vaccines14040330

Figure Lengend Snippet: Toxicity screening of CARDS mutants and purity analysis of recombinant proteins. ( A ) Dose–response curves of TNF-α secretion from RAW264.7 cells treated with CARDS-WT and various mutants, used for screening attenuated mutants. ( B – H ) HPLC chromatograms of recombinant proteins on TSKgel G2000SWXL column. Purity values are indicated above the main peaks: ( B ) CARDS-E132A (78.88%), ( C ) CARDS-E132Q (73.47%), ( D ) CARDS-H36A (86.99%), ( E ) CARDS-R10A (82.72%), ( F ) P1 (87.65%), ( G ) P40/90 (96.76%), ( H ) CARDS-WT (91.77%).

Article Snippet: The coding sequences for P1 (GenBank I D: X06871 ), P40 (GenBank ID: AF125204 ), P90 (GenBank ID: AF125205 ), and CARDS-WT (GenBank ID: AY189944 ) were retrieved from the NCBI database based on the Mycoplasma pneumoniae standard strain ATCC M129.

Techniques: Recombinant

Journal: Vaccines

Article Title: Three-Component Subunit Vaccine Induces Protective Immunity Against Mycoplasma pneumoniae in Mice

doi: 10.3390/vaccines14040330

Figure Lengend Snippet: Formulation, immunization schedule, and antigen-specific IgG antibody responses of the three-component M. pneumoniae vaccine. ( A ) Formulation scheme showing the 6 experimental groups, their respective vaccine and adjuvant combinations. ( B ) Immunization schedule showing time points for immunizations (days 0, 14, 28) and serum collection (days 13, 27, 41). ( C – F ) Serum titers (log10) of specific IgG antibodies against P1, P40/90, CARDS-WT, and CARDS-Mut following single antigen immunizations (days 0, 14, 28) and serum collection (days 13, 27, 41). The saline group served as the negative control. ( G – J ) Serum antibody titers (days 41) against P1, P40/90, CARDS-WT, and CARDS-Mut in mice immunized according to the groups shown in ( A ). ( K ) Comparison of antibody titers (days 41) between the MPtriVa-D group (containing CARDS-WT with dual adjuvant) and the MPtriVb-D group (containing CARDS-Mut with dual adjuvant). ( L ) Comparison of antibody titers (days 41) between the MPtriVb-S (single adjuvant) and MPtriVb-D (dual adjuvant) groups. Data are presented as mean ± SD. p > 0.05 (not significant); *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: The coding sequences for P1 (GenBank I D: X06871 ), P40 (GenBank ID: AF125204 ), P90 (GenBank ID: AF125205 ), and CARDS-WT (GenBank ID: AY189944 ) were retrieved from the NCBI database based on the Mycoplasma pneumoniae standard strain ATCC M129.

Techniques: Formulation, Adjuvant, Saline, Negative Control, Comparison

Journal: Biomolecular NMR assignments

Article Title: Backbone NMR resonance assignments for the C terminal domain of the Streptococcus mutans adhesin P1

doi: 10.1007/s12104-023-10158-y

Figure Lengend Snippet: (a) Annotated 2D 1H,15N TROSY spectrum of Adhesin P1 C3 domain (BMRB 52097) collected in an 800 MHz spectrometer at 25 °C in phosphate buffer pH 6. Resonance assignments are shown with black labels; (b) Alphafold structural model for the new, longer C3 construct with the amino acid residues that are currently assigned shown in blue; (c) C3 domain structure (PDB 3QE5) with the residues assigned using the previous, shorter C3 construct (BMRB 27935) shown in red; (d) comparison of 2D 1H,15N TROSY spectra for the previous, shorter C3 construct (red) and the new, longer construct for the AlphaFold-predicted C3 domain (blue). Disordered, poorly resolved resonances observed for the prior C3 construct are now well resolved for the new C3 construct by addition of the seven C-terminal amino acids.

Article Snippet: The DNA sequence coding for the C3 domain of adhesin P1 (residues 1328–1490; Uniprot accession number P23504) was synthesized, with codon optimization for E. coli , by Genescript and inserted into pET21a(+) plasmid to yield the pET21a-C3-His 6 plasmid with a C-terminal His tag.

Techniques: Construct, Comparison