Journal: Frontiers in Veterinary Science

Article Title: Construction and immunogenicity evaluation of a bivalent nanoparticle based on mi3 displaying porcine circovirus type 2 and type 3 capsid proteins

doi: 10.3389/fvets.2026.1862938

Figure Lengend Snippet: Construction of engineered E. coli strains and protein purification. (A) PCR identification of positive E. coli clones. Lanes 1–3 correspond to BL21-pET28-ST-PCV2 Cap, BL21-pET28-ST-PCV3 Cap, and BL21-pET28-SC-mi3, respectively. (B–D) Expression and purification of recombinant proteins ST-PCV2 Cap, ST-PCV3 Cap, and SC-mi3, respectively. Lanes 1–6 represent whole bacterial lysate, pellet after sonication, supernatant after sonication, Ni column flow-through, wash fraction, and purified protein, respectively. (E) Western blot validation of antigen proteins. Lanes 1, 2 correspond to ST-PCV2 Cap and ST-PCV3 Cap, respectively. (F) Endotoxin removal from recombinant protein solutions. Color intensity represents endotoxin content; numbers indicate endotoxin content (EU/mL).

Article Snippet: The pET28a(+) plasmid for recombinant protein expression and BL21(DE3) E. coli competent cells were purchased from Sangon Biotech (Shanghai) Co., Ltd.

Techniques: Protein Purification, Clone Assay, Expressing, Purification, Recombinant, Sonication, Western Blot, Biomarker Discovery

Journal: bioRxiv

Article Title: Cryo-EM structure of MutL-activated MutH

doi: 10.64898/2026.04.21.719898

Figure Lengend Snippet: (A) Structure of the MutL-MutH-DNA complex. MutL LN40 is coloured in light and dark green, MutH in purple, and the DNA in orange. (B) Structure of MutL LN40 bound to a 5′ extended single stranded DNA (PDB ID: 7P8V). In this structure, the single-stranded DNA overhang is located in the same groove as MutH in the MutL-MutH-DNA complex, indicting that MutL LN40 cannot bind MutH and DNA simultaneously. (C) Superposition of three MutL LN40 structures in different functional states: apo, ssDNA-bound, and MutH-bound. The changes in the groove between the two monomers are highlighted by helices 265–281 and 313–331. The width of the groove ranges from 14 Å in the ssDNA-bound structure (blue, PDB ID: 7P8V), to 21 Å in the apo structure (orange, PDB ID: 1B63), to 28 Å in the MutH-bound structure (green, this work. (D) Conservation analysis of MutH from H. influenzae and E. coli mapped onto the E. coli MutH structure. Highly conserved residues are shown in magenta, and lesser conserved residues in blue. The strong conservation of the DNA-binding site supports the use of the available H. influenzae MutH-DNA structure (PDB ID: 2AOR) as a structural reference for comparison with our E. coli MutL-bound MutH complex.

Article Snippet: All proteins were expressed in E. coli BL21(DE3) cells grown in 2x YT media (Formedium, UK) at 30 °C.

Techniques: Functional Assay, Binding Assay, Comparison