Journal: Poultry Science

Article Title: Construction and modification of a low-copy plasmid-based infectious clone for GI-19 genotype IBV via Red/ET recombineering: A simplified and efficient reverse genetics system for co ronavirus

doi: 10.1016/j.psj.2026.106881

Figure Lengend Snippet: Validation of recombinant plasmids by restriction enzyme analysis and functional selection screening. (A) Restriction enzyme digestion of intermediate recombinant plasmids was performed using XhoI, XbaI , and KpnI . The observed banding patterns were consistent with the predictions generated by SnapGene software. Lanes 1–3 show the predicted digestion patterns of p15A-CmR-D90, p15A-CmR-D90-Δ5a/AmpR-ccdB, and p15A-CmR-D90-Δ5a/EGFP. Lanes 4–5, 6–7, and 8–9 correspond to the experimental digestion results of these respective plasmids. M: TaKaRa 1 kb DNA Ladder. All digested products were resolved by electrophoresis on 1% agarose gels and visualized with ethidium bromide staining. (B) Functional verification of the selection marker gene. The intermediate recombinant plasmid p15A-CmR-D90-Δ5a/AmpR-ccdB was transformed into two E. coli strains: GBred (CcdB-sensitive) and GBred- gyrA462 (CcdB-resistant). On LB agar plates containing ampicillin and chloramphenicol, only GBred- gyrA462 survived, confirming the toxicity of CcdB and functionality of the selection cassette. In the second recombination step, replacement of the AmpR-ccdB cassette by the target Δ5a/EGFP fragment yielded the final recombinant plasmid p15A-CmR-D90-Δ5a/EGFP. Upon counter-selection with CcdB, only host E. coli GBred harboring the correctly recombined plasmid survived on chloramphenicol-containing LB plates, while non-recombinant bacteria were eliminated.

Article Snippet: M: TaKaRa 1 kb DNA Ladder.

Techniques: Biomarker Discovery, Recombinant, Functional Assay, Selection, Generated, Software, Electrophoresis, Staining, Marker, Plasmid Preparation, Transformation Assay, Bacteria

Journal: International Journal of Pharmaceutics: X

Article Title: Nanomedicines for DNA and interference RNA co-delivery: Combined gene therapy for Fabry disease

doi: 10.1016/j.ijpx.2026.100555

Figure Lengend Snippet: Schematic representation of combined gene therapy for FD. α-Gal A: α-Galactosidase A. pDNA: Plasmid DNA. Gb3S: Globotriaosylceramide synthase. siRNA: Small interference RNA. FD: Fabry disease. gSRT: Genetic substrate reduction therapy. Gb3: Globotriaosylceramide.

Article Snippet: Agarose, deoxyribonuclease I (DNAse I) and sodium dodecyl sulfate (SDS) were purchased from Sigma-Aldrich, GelRedTM from Biotium (Fremont, CA, USA) and GeneRuler Ultra Low Range DNA Ladder and 1 Kb pDNA ladder from Gibco (Thermo Fisher Scientific, MA, USA).

Techniques: Plasmid Preparation

Journal: International Journal of Pharmaceutics: X

Article Title: Nanomedicines for DNA and interference RNA co-delivery: Combined gene therapy for Fabry disease

doi: 10.1016/j.ijpx.2026.100555

Figure Lengend Snippet: Binding, protection and release capacity of co-delivery vectors with SLNs containing DOTAP or DOTAP/MC3. In all gels, lane 1 corresponds to 1 Kb pDNA ladder. Lane 2 to naked pCMV6-AC-αGLA, lane 3 to naked pCMV6-AC-αGLA + SDS, lane 4 to naked siRNA, lane 5 to siRNA + SDS, lane 6 to siRNA-GN + SDS, lane 11 to naked pR-M10-αGal A and lane 12 to Naked siRNA + DNase + SDS. ( A ) DxP3GNSLNEE co-delivery vectors: (7) DxP3GN5-ET. (8) DxP3GN2.5-ET. (9) DxP3GN5-EMC3. (10) DxP3GN2.5-EMC3. (13) DxP3GN5-ET vector + DNase + SDS. (14) DxP3GN2.5-ET vector + DNase + SDS. (15) DxP3GN5-EMC3 + DNase + SDS. (16) DxP3GN2.5-EMC3 + DNase + SDS. (17) DxP3GN5-ET + SDS. (18) DxP3GN2.5-ET + SDS. (19) DxP3GN5-EMC3 + SDS. (20) DxP3GN2.5-EMC3 + SDS. (B) HAP3GNSLNEE co-delivery vectors: (7) HAP3GN5-ET. (8). HAP3GN2.5-ET. (9) HAP3GN5-EMC3. (10) HAP3GN2.5-EMC3. (13) HAP3GN5-ET + DNase + SDS. (14) HAP3GN2.5-ET + DNase + SDS. (15) HAP3GN5-EMC3 + DNase + SDS. (16) HAP3GN2.5-EMC3 + DNase + SDS. (17) HAP3GN5-ET + SDS. (18) HAP3GN2.5-ET + SDS. (19) HAP3GN5-EMC3 + SDS. (20) HAP3GN2.5-EMC3 + SDS. ( C ) DxP2GNSLNEE co-delivery vectors: (7) DxP2GN5-ET. (8). DxP2GN2.5-ET. (9) DxP2GN5-EMC3. (10) DxP2GN2.5-EMC3. (13) DxP3GN5-ET + DNase + SDS. (14) DxP3GN2.5-ET + DNase + SDS. (15) DxP2GN5-EMC3 + DNase + SDS. (16) DxP2GN2.5-EMC3 + DNase + SDS. (17) DxP2GN5-ET + SDS. (18) DxP2GN2.5-ET + SDS. (19) DxP2GN5-EMC3 + SDS. (20) DxP2GN2.5-EMC3 + SDS. ( D ) HAP2GNSLEE co-delivery vectors: (7) HAP2GN5-ET. (8). HAP2GN2.5-ET. (9) HAP2GN5-EMC3. (10) HAP2GN2.5-EMC3. (13) HAP2GN5-ET + DNase + SDS. (14) HAP2GN2.5-ET + DNase + SDS. (15) HAP2GN5-EMC3 + DNase + SDS. (16) HAP2GN2.5-EMC3 + DNase + SDS. (17) HAP2GN5-ET + SDS. (18) HAP2GN2.5-ET + SDS. (19) HAP2GN5-EMC3 + SDS. (20) HAP2GN2.5-EMC3 + SDS.

Article Snippet: Agarose, deoxyribonuclease I (DNAse I) and sodium dodecyl sulfate (SDS) were purchased from Sigma-Aldrich, GelRedTM from Biotium (Fremont, CA, USA) and GeneRuler Ultra Low Range DNA Ladder and 1 Kb pDNA ladder from Gibco (Thermo Fisher Scientific, MA, USA).

Techniques: Binding Assay, Plasmid Preparation