bl21  (New England Biolabs)

Journal: mBio

Article Title: A Multiprotein DNA Translocation Complex Directs Intramycelial Plasmid Spreading during Streptomyces Conjugation

doi: 10.1128/mBio.02559-14

Figure Lengend Snippet: SpdB2-His promotes spheroplast formation by T7 lysozyme and forms unstable pores in planar lipid bilayers. (A) When expression of SpdB2-His was induced in BL21/pLysS, spheroplast (arrows) were formed. In contrast, spheroplasts were not observed in the absence of spdB2 (B) or when expressing spdB2 in BL21 lacking the T7 amidase (C). (D) When purified SpdB2-His was added to the cis side of a lipid bilayer, SpdB2-His inserted into the planar lipid bilayer, showing flickering pore structures. (E) The corresponding all-points conductance level histogram shows that there is no discrete conductance state of the open pore indicated by the long tail in the histogram.

Article Snippet: The resulting expression plasmid pGMT7-spdB2 was used to transform the different E. coli strains [BL21(DE3), BL21(DE3)/pLysS, Lemo21(DE3) (NEB)].

Techniques: Expressing, Purification

Journal: Microbial Cell Factories

Article Title: Investigation of the methylerythritol 4-phosphate pathway for microbial terpenoid production through metabolic control analysis

doi: 10.1186/s12934-019-1235-5

Figure Lengend Snippet: Specific efflux rates of MEP pathway intermediates calculated from their measured concentrations in the supernatant of cultures of the E. coli Bl21 dxs RBS library expressing isp S and idi from pCOLA::IspS-idi ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\overline{\varvec{x}}$$\end{document} x ¯ ± SD, n = 3). The cultures were induced at OD 600 = 0.1 and the supernatant was sampled at OD 600 ≈ 0.5. The export of MEP pathway intermediates occurred after the induction of isp S expression in the dxs expression library. a Metabolite concentration in the supernatant of the cultures. b DXP and MEcPP efflux rates as a function of dxs expression relative to wild-type levels. Linear fits through zero and second-degree polynomial fits through zero are shown for DXP and MEcPP, respectively in ( a ) and ( b ). c Logarithmic plot of MEcPP efflux rate as a function of dxs expression. The data were fitted to a linear function with a slope of 2.0. The red marked data point was excluded from the fit. d The correlation of MEcPP efflux rate with its intracellular concentration, linear fitted through zero. e Combined efflux of DXP, MEcPP and isoprene in dependents of dxs expression, linear fitted

Article Snippet: Bacterial strains and culture conditions Standard cloning and metabolic engineering were carried out using E. coli strains 10β and BL21 (DE3), respectively (both supplied by New England Biolabs, Ipswich, MA, USA).

Techniques: Expressing, Concentration Assay

Journal: Avicenna Journal of Medical Biotechnology

Article Title: Molecular characterization and Functional Analysis of the PilQ380-706: a Novel Secretin Domain in Pseudomonas aeruginosa

doi:

Figure Lengend Snippet: Western blot analysis of the expressed r-PilQ 380-706 protein in E. coli BL21. After running the SDS-PAGE, the protein transferred onto PVDF membrane and detected with HRP-conjugated goat anti-rabbit IgG. (lane 1) total cell lysate of non-induced bacteria; (lane 2) total cell lysate of bacteria after 4 hr induction; (lane 3 and 4) purified r-PilQ 380-706 by Ni 2+ -affinity chromatography.

Article Snippet: Bacterial strains, plasmids, and media Escherichia coli (E. coli ) strains Top10F and BL21 (DE3) were used as preservation and expression hosts.

Techniques: Western Blot, SDS Page, Purification, Affinity Chromatography

Journal: bioRxiv

Article Title: Pyocin S5 import into Pseudomonas aeruginosa reveals a generic mode of bacteriocin transport

doi: 10.1101/856047

Figure Lengend Snippet: FptA and TonB1 constitute the minimal system for PyoS5-susceptibility in E. coli . (A) Susceptibility to PyoS5 (3 and 60 μM) was assessed for P. aeruginosa YHP17, (B) for E. coli BL21 (DE3) expressing FptA, (C) for E. coli BL21 (DE3) expressing TonB-B1, and (D) for E. coli BL21 (DE3) expressing FptA and TonB-B1. Zones of clearance are observed in all E. coli samples for the ColBPyoS5 (13 μM) control (B-D) and for both concentrations of PyoS5 in E. coli expressing FptA and TonB-B1 (D). In the P. aeruginosa control clearance zones were observed for PyoS5 (A).

Article Snippet: Chemically competent E. coli NEB5α and BL21 (DE3) were purchased from NEB.

Techniques: Expressing

Journal: bioRxiv

Article Title: Pyocin S5 import into Pseudomonas aeruginosa reveals a generic mode of bacteriocin transport

doi: 10.1101/856047

Figure Lengend Snippet: FpvAI and TonB1 constitute the minimal system for PyoS2- and PyoS4-susceptibility. Susceptibility to 1 μM PyoS2, PyoS4, PyoS5 and ColBPyoS5 was assessed for (A) P. aeruginosa YHP17, (B) E. coli BL21 (DE3) expressing FpvAI, (C) E. coli BL21(DE3) expressing TonB-B1, and (D) E. coli BL21 (DE3) expressing FpvAI and TonB-B1. Zones of clearance were observed for all E. coli conditions tested when ColBPyoS5 was applied (B-D). Similarly, E. coli expressing FpvAI and TonB-B1 was susceptible to both PyoS2 and PyoS4, as was the P. aeruginosa control (A+D). Zones of clearance for PyoS5 were only observed in P. aeruginosa .

Article Snippet: Chemically competent E. coli NEB5α and BL21 (DE3) were purchased from NEB.

Techniques: Expressing