Phenotypic and Genetic Characterization of Carbapenemase and ESBLs Producing Gram-negative Bacteria (GNB) Isolated from Patients with Cystic Fibrosis in Tehran Tospitals

Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we extract, purify and reconstitute them into artificial planar membranes. To avoid this time-consuming procedure, here we show a robust approach using fusion of native outer membrane vesicles (OMV) into planar lipid bilayer which moreover allows also to some extend the characterization of membrane protein channels in their native environment. Two major membrane channels from Escherichia coli, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion revealed surprisingly single or only few channel activities. The asymmetry of the OMV´s translates after fusion into the lipid membrane with the LPS dominantly present at the side of OMV addition. Compared to conventional reconstitution methods, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution. The addition of Enrofloxacin on the LPS side yields somewhat higher association (kon) and lower dissociation (koff) rates compared to LPS-free reconstitution. We conclude that using outer membrane vesicles is a fast and easy approach for functional and structural studies of membrane channels in the native membrane.

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Discovery and Characterization of the Antimetabolite Action of Thioacetamide-Linked 1,2,3-Triazoles as Disruptors of Cysteine Biosynthesis in Gram-Negative Bacteria

ACS Infectious Diseases ◽

10.1021/acsinfecdis.9b00406 ◽

2019 ◽

Vol 6 (3) ◽

pp. 467-478 ◽

Cited By ~ 1

Author(s):

Miranda J. Wallace ◽

Suresh Dharuman ◽

Dinesh M. Fernando ◽

Stephanie M. Reeve ◽

Clifford T. Gee ◽

...

Keyword(s):

Gram Negative Bacteria ◽

Cysteine Biosynthesis ◽

Gram Negative

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Phenotypic and genetic characterization of a patient with a de novo interstitial 14q24.1q24.3 deletion

Molecular Cytogenetics ◽

10.1186/1755-8166-7-49 ◽

2014 ◽

Vol 7 (1) ◽

pp. 49 ◽

Cited By ~ 2

Author(s):

Elisa Tassano ◽

Andrea Accogli ◽

Serena Panigada ◽

Patrizia Ronchetto ◽

Cristina Cuoco ◽

...

Keyword(s):

De Novo ◽

Genetic Characterization ◽

Phenotypic And Genetic Characterization

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Susceptibility of Gram-negative bacteria to β-lactam antibiotics and rapid characterization of β-lactamase activity

Current Microbiology ◽

10.1007/bf02601720 ◽

1979 ◽

Vol 2 (4) ◽

pp. 239-244 ◽

Cited By ~ 1

Author(s):

David L. Gibbs ◽

Clyde Thornsberry

Keyword(s):

Gram Negative Bacteria ◽

Gram Negative ◽

Rapid Characterization

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Protein expression and extraction of hard-to-produce proteins in the periplasmic space of Escherichia coli v1

10.17504/protocols.io.bxxxpppn ◽

2021 ◽

Author(s):

Cristina Hernandez Rollan ◽

Kristoffer Bach Falkenberg ◽

Maja Rennig ◽

Andreas Birk Bertelsen ◽

Morten Norholm

Keyword(s):

Protein Production ◽

Expression System ◽

Gram Negative Bacteria ◽

Gram Negative ◽

E Coli ◽

New Family ◽

Lytic Polysaccharide Monooxygenases ◽

Strain Bl21 ◽

Polysaccharide Monooxygenases

E. coli is a gram-negative bacteria used mainly in academia and in some industrial scenarios, as a protein production workhorse. This is due to its ease of manipulation and the range of genetic tools available. This protocol describes how to express proteins in the periplasm E. coli with the strain BL21 (DE3) using a T7 expression system. Specifically, it describes a series of steps and tips to express "hard-to-express" proteins in E. coli, as for instance, LPMOs. The protocol is adapted from Hemsworth, G. R., Henrissat, B., Davies, G. J., and Walton, P. H. (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat. Chem. Biol.10, 122–126. .

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SKRENING BAKTERI TOLERAN PESTISIDA DENGAN BAHAN AKTIF KLORANTRANILIPROL ASAL TANAH PERTANIAN BATURITI TABANAN BALI

Jurnal Biologi Udayana ◽

10.24843/jbiounud.2017.vol21.i01.p01 ◽

2017 ◽

Vol 21 (1) ◽

pp. 1

Author(s):

NI KADEK WIWIK SINTA DEWI ◽

IDA BAGUS GEDE DARMAYASA ◽

I KETUT SUNDRA

Keyword(s):

Large Scale ◽

Chemical Compounds ◽

Mineral Medium ◽

Gram Negative Bacteria ◽

Soil Mineral ◽

Gram Negative ◽

Gram Positive Bacteria ◽

Numerous Problem ◽

Stage 1

In Indonesia agriculture practice often used the large scale pesticide application such as insecticide, herbicide, and fungicide. The wide use of toxic pesticide has created numerous problem in increasing environtmental hazard to human and to other animals. Many of soil bacteria had important role to degrading chemical compounds into simpler compounds as a bioremediation agent. The aim of this study was to screen the Chlorantraniliprole tolerant bacteria using soil mineral medium with Prevathon pesticide addition, also teo identificate the species of bacteria. This research was conducted at Microbiology Laboratory, Faculty of Mathematics and Natural Sciences, Udayana University. The research was done in three analysis, (1) bacteria test on Prevathon pesticide addition to mineral medium treatment, (2) characterization of bacteria, (3) Identification of pesticide tolerant bacteria with BD BBL Crystal Enteric/Non FermenterID Kit. The results showed that mineral medium with the addition of pesticides Prevathon treatment able to provide a significant different effect on the enrichment stage 1, stage 2 and stage 3 (P<0.05), there was 5 isolates pesticide tolerant bacteria that isolated from Baturiti Tabanan cultivated soil that was BSP 1, BSP 2, BSP 3 known as gram negative bacteria, and BSP 4, BSP 5 known as Gram positive bacteria, pesticide tolerant bacteria identified as Serratia marcescens which is a Gram negative bacteria group and may cause pathogenic.

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