e coli wm3064  (Addgene inc)

Journal: iScience

Article Title: Origin of replication discovery for environmentally isolated Pantoea strain enables expression of heterologous proteins, pathways and products

doi: 10.1016/j.isci.2026.115031

Figure Lengend Snippet: Magic pool BarSeq results for all origin sequences tested The y axis denotes the names of the origins of replication, with predicted origins in green, and origins found in literature in black. Colors in each time point, or source, square represent the fraction of barcode counts associated with that origin versus the total fraction of barcodes in that sample (see legend). The number present in each box represents the total number of unique barcodes detected for that particular origin of replication at that time point, or for that source. (A) Plate-based sources for the magic pool conjugations with the 3 hosts. “Plate” is indicative of cells harvested directly from an agar plate, while “Outgrowth” is indicative of an overnight of growth in liquid media after scraping colonies off of an agar plate. (B) Liquid-based time points for magic pool conjugations with the 3 hosts. Time points 1, 2, 3 and 4 represent 24, 48, 72, and 96 h respectively. Note that the recipient strain E. coli BW25113 is part of the recipient set along with Pantoea MT58 and Brevundimonas sp , and is different from the conjugation strain E. coli WM3064, which was used as the donor. Note also that the 3845 hit for Brevundimonas sp. was analyzed due to a single bar code enrichment and represents an artifact rather than a real candidate.

Article Snippet: Competent cells of E. coli TransforMax EC100D pir -116 cloning strain and the E. coli WM3064 conjugation donor strain were purchased from Lucigen (Biosearch Technologies, Alexandria, MN).

Techniques: Conjugation Assay

Journal: Foods

Article Title: Identification of a TonB-Dependent Siderophore Receptor as a Novel Anti-Biofilm Target and Virtual Screening for Its Inhibitor in Pseudomonas fluorescens PF08

doi: 10.3390/foods14030531

Figure Lengend Snippet: Bacterial strains and plasmids used in this work.

Article Snippet: E. coli WM3064 , Host for pir -dependent plasmids and donor strain for conjugation; RP4 ( tra ) in chromosome; Δ dapA , Originally from BioVector NTCC.

Techniques: Conjugation Assay, Expressing, Plasmid Preparation

Journal: Microbial cell factories

Article Title: Engineering osmolysis susceptibility in Cupriavidus necator and Escherichia coli for recovery of intracellular products.

doi: 10.1186/s12934-023-02064-8

Figure Lengend Snippet: Fig. 5 Application of mechanosensitive knockout for osmolysis in E. coli BL21. A Percent cell lysis following growth in LB with 4% NaCl of BL21 (blue), BL21 ΔmscL (red), and BL21 ΔmscL ΔmscS (yellow) in three different media: commercial B-PER™ Bacterial Protein Extraction Reagent; a 4% NaCl(aq) isotonic solution; and distilled water (diH2O). Cell lysis is determined as the fraction of RFP recovered in supernatant compared to total RFP content, as it was in C. necator, with concentration measured by fluorescence intensity (n = 6). B Growth rate (red diamonds) and osmolysis efficiency (blue circles) of BL21 ΔmscL ΔmscS in LB with various (final) NaCl concentrations (n = 3). C SDS-PAGE gel of various fractions of RFP-expressing BL21 and BL21 ΔmscL ΔmscS. NE: cells not expressing RFP, WC: whole-cell fraction, S: post-osmolysis supernatant fraction, P: post-osmolysis cell pellet. For each data point in (A) and (B), mean ± SD is displayed

Article Snippet: All C. necator strains listed in Table 1 were transformed with the red fluorescent protein (RFP) expression plasmid pBADTrfp via conjugation using E. coli WM3064 as a donor, following the protocol from Windhorst et al. [31]. pBADTrfp was a gift from Nathan Hillson (Addgene plasmid # 99382; http:// n2t. net/ addge ne: 99382; RRID:Addgene_99382) [29]. pBADTrfp was transformed into chemically competent WM3064 cells using heat shock and positive clones were selected for on LB-Agar plates containing DAP and kanamycin (50 μg/ mL) following a 1-h outgrowth in SOC medium.

Techniques: Knock-Out, Lysis, Protein Extraction, Concentration Assay, Fluorescence, SDS Page, Expressing