Journal: bioRxiv

Article Title: Native plasmid-encoded mercury resistance genes are functional and demonstrate natural transformation in environmental bacterial isolates

doi: 10.1101/764415

Figure Lengend Snippet: Plasmid map of p5343_UC57. The p5343 part (depicted in blue) encodes Mercuric ion reductase (MerA), Mercuric ion uptake protein (MerF), Hyp: Hypothetical protein (Hyp2), Mercuric transport protein (MerP), Mercuric transport protein (MerT), Regulator of Mercury resistance genes (MerR), Mobilization protein A (MobA), Mobilization protein C (MobC), Plasmid replication protein (RepA), Helix-turn-helix domain protein (HTH), RelE toxin (RelE), while the pUC57 part (depicted in grey) codes for promotor sequence for Ampicillin resistance gene (AmpR Promotor), Ampicillin resistance marker (AmpR), origin of replication, Lac promotor, LEU2 selection marker (Leu2) and elements ensuring plasmid maintenance (CEN/ARS).

Article Snippet: To propagate the synthesized DNA in the model bacterium E. coli , p5343 was cloned into the vector pUC57 resulting in plasmid p5343_UC57 (plasmid and sequence available via Addgene 126645).

Techniques: Plasmid Preparation, Sequencing, Marker, Selection

Journal: bioRxiv

Article Title: Native plasmid-encoded mercury resistance genes are functional and demonstrate natural transformation in environmental bacterial isolates

doi: 10.1101/764415

Figure Lengend Snippet: Improved mercury resistance in Escherichia coli DH10B containing p5343_UC57 (blue) versus only the empty vector pUC57 (green), grown in the presence of varying mercuric chloride concentrations.

Article Snippet: To propagate the synthesized DNA in the model bacterium E. coli , p5343 was cloned into the vector pUC57 resulting in plasmid p5343_UC57 (plasmid and sequence available via Addgene 126645).

Techniques: Plasmid Preparation

Journal: bioRxiv

Article Title: Native plasmid-encoded mercury resistance genes are functional and demonstrate natural transformation in environmental bacterial isolates

doi: 10.1101/764415

Figure Lengend Snippet: Results of the HGT assay used to screen environmental isolates for uptake of a plasmid-encoded trait via natural transformation. The 58 bacterial isolates are depicted on a 16S rRNA sequence-based phylogenetic tree (constructed using Maximum Likelihood). The colored bars indicate different bacterial classes, namely, orange is Actinobacteria, red is Cytophagia (Bacteriodetes phylum), yellow is Alphaproteobacteria, peach is Betaproteobacteria and pink is Gammaproteobacteria. The heat map plots optical density in the presence of p5343_UC57 compared to pUC57 in the presence of different mercury chloride concentrations normalized by the no mercury chloride control. Only the strains showing improved plasmid-based growth in the presence of mercury chloride were subjected to merA based colony PCR. The strains positive for colony PCR are indicated in black circles while those negative are depicted in grey circles. The genus of the strains positive for natural transformation as depicted on the figure are Arthrobacter (Ar), Dermacoccus (De), Acidovorax (Ax), Cupriavidus (Cu), Acinetobacter (Ac) and Pseudomonas (Ps). Figure constructed using ITOL 46 .

Article Snippet: To propagate the synthesized DNA in the model bacterium E. coli , p5343 was cloned into the vector pUC57 resulting in plasmid p5343_UC57 (plasmid and sequence available via Addgene 126645).

Techniques: Plasmid Preparation, Transformation Assay, Sequencing, Construct

Journal: bioRxiv

Article Title: Native plasmid-encoded mercury resistance genes are functional and demonstrate natural transformation in environmental bacterial isolates

doi: 10.1101/764415

Figure Lengend Snippet: Confirmation of HGT in strains 5, 12 and DH10B. Growth curves from second growth regime in the presence of varying mercury chloride concentrations are depicted when strains 5, 12 and DH10B were naturally transformed with plasmid p5343_UC57 (blue) and pUC57 (green). Assay design provided in Figure S2.

Article Snippet: To propagate the synthesized DNA in the model bacterium E. coli , p5343 was cloned into the vector pUC57 resulting in plasmid p5343_UC57 (plasmid and sequence available via Addgene 126645).

Techniques: Transformation Assay, Plasmid Preparation