Journal: npj Antimicrobials and Resistance

Article Title: A conjugal gene drive-like system efficiently suppresses antibiotic resistance in a bacterial population

doi: 10.1038/s44259-026-00181-z

Figure Lengend Snippet: a Schematic of p Pro-MobV and p ETas plasmids. Left, donor E. coli EPI300 containing p Pro-MobV includes the following components: medium copy ori pBBR1, conjugative origin of transfer ( oriT) , arabinose-inducible λRed and Cas9, sgRNA targeting bla (Amp R ) flanked with HAs, chloramphenicol resistance (Cm R ) and Gm R resistance genes. Conjugative machinery, genes required for conjugation are derived from the IncP RK2 conjugative system. Right, E. coli MG1655 recipient cells carrying p ETas plasmid. b Scheme of conjugation using donor and recipient cells, selection of transconjugants on triple antibiotics plate (Amp + Sm + Cm), overnight culture with and without arabinose and plating on Amp or Sm plates. c Recovery of CFU on Amp plates (solid blue and red dots) and Sm plates (open blue and red dots) following Control and Pro-AG experiments. d DNA sequence analysis of plasmids isolated from single colonies from Sm plates after Pro-AG events. e Scheme depicting HBD-mediated deletion of bla locus via recombination between the directly repeated promoters driving expression of the Amp R and Sm R genes. Data were plotted as the mean ± SEM, representing three independent experiments performed in triplicate and analyzed by Student’s t- test. NS not significant ( P > 0.05); *** P < 0.001.

Article Snippet: The ligated product was electroporated into TransforMax EPI300 Electrocompetent E.coli (Lucigen) as described earlier, and cells were resuspended in super-optimal broth with catabolite repression media.

Techniques: Conjugation Assay, Derivative Assay, Plasmid Preparation, Selection, Control, Sequencing, Isolation, Expressing

Journal: npj Antimicrobials and Resistance

Article Title: A conjugal gene drive-like system efficiently suppresses antibiotic resistance in a bacterial population

doi: 10.1038/s44259-026-00181-z

Figure Lengend Snippet: a Donor bacteria EPI300 containing p Pro-MobV constitutes medium copy ori pBBR1, conjugative origin of transfer ( oriT) , arabinose-inducible λRed and Cas9, sgRNA targeting bla (Amp R ) flanked with HAs, chloramphenicol (Cm R ) and gentamicin (Gm R ) resistance gene. Conjugative machinery, genes required for conjugation and E. coli MG1655 Δ recA recipient cells carrying p ETas plasmid. b Tabulation of editing outcomes of CFU recovered on Amp plates and Sm plates following Control and Pro-AG experiments. c DNA sequence analysis of plasmids isolated from single colonies from Sm plates after Pro-AG events. d Schematic of plasmids used to compare HBD events, p Pro-AG , p Pro-AG-ΔCas9 (Cas9 deleted version of p Pro-AG ), p Pro-AG-ΔλRed (λRed deleted version of p Pro-AG ), and target E. coli MG1655 WT or isogenic Δ recA cells carrying p ETas plasmid. e Comparison of CFU following Control and Pro-AG using p Pro-AG , p Pro-AG-ΔCas9 and p Pro-AG-ΔλRed donor plasmids in WT or Δ recA receiver cells. f DNA sequence analysis of target plasmids isolated from single colonies from Sm plates after Pro-AG events. Data were plotted as the mean ± SEM, representing three independent experiments performed in triplicate and analyzed by Student’s t -test. NS not significant ( P > 0.05); **** P < 0.0001.

Article Snippet: The ligated product was electroporated into TransforMax EPI300 Electrocompetent E.coli (Lucigen) as described earlier, and cells were resuspended in super-optimal broth with catabolite repression media.

Techniques: Bacteria, Conjugation Assay, Plasmid Preparation, Control, Sequencing, Isolation, Comparison

Journal: Bio-protocol

Article Title: Cloning a Chloroplast Genome in Saccharomyces cerevisiae and Escherichia coli

doi: 10.21769/BioProtoc.5162

Figure Lengend Snippet: A) DNA isolated from yeast transformants that screen positively is electroporated into E. coli for further analysis. B) A 2% agarose gel where 2 μL of the multiplex PCR reactions were visualized. All E. coli transformants demonstrate the expected multiplex banding pattern, suggesting that the whole chloroplast genome has been successfully transformed. The positive control consists of total DNA isolated from P. tricornutum and the negative control has no DNA added.

Article Snippet: E. coli TransforMax EPI300 cells (LGC Biosearch Technologies, Lucigen, catalog number: EC300110) 3.

Techniques: Isolation, Agarose Gel Electrophoresis, Multiplex Assay, Transformation Assay, Positive Control, Negative Control

Journal: Microbial Cell Factories

Article Title: Uncovering the genetic basis of antiviral polyketide limocrocin biosynthesis through heterologous expression

doi: 10.1186/s12934-024-02621-9

Figure Lengend Snippet: Testing of biological activity and cytotoxicity of limocrocins. A antimicrobial activities of limocrocins against selected microorganisms. B half maximal inhibitory concentrations (IC 50 ) ± standard deviation against Chinese hamster ovary cells (CHO-K1)

Article Snippet: The ligated DNA was packaged into phage particles and then used to infect E. coli EPI300 cells (TransforMax™ EPI300™-T1 R Electrocompetent E. coli Kit, Biosearch Technologies).

Techniques: Activity Assay, Standard Deviation

Journal: Microbial Cell Factories

Article Title: Uncovering the genetic basis of antiviral polyketide limocrocin biosynthesis through heterologous expression

doi: 10.1186/s12934-024-02621-9

Figure Lengend Snippet: Plasmids and bacterial strains used in this work

Article Snippet: The ligated DNA was packaged into phage particles and then used to infect E. coli EPI300 cells (TransforMax™ EPI300™-T1 R Electrocompetent E. coli Kit, Biosearch Technologies).

Techniques: Plasmid Preparation, Expressing, Clone Assay, Derivative Assay, Cosmid DNA