Cloning and characterization of an aminoglycoside 6'-N-acetyltransferase gene from Citrobacter freundii which confers an altered resistance profile.

1997 ◽  
Vol 41 (11) ◽  
pp. 2439-2447 ◽  
Author(s):  
H Y Wu ◽  
G H Miller ◽  
M G Blanco ◽  
R S Hare ◽  
K J Shaw
Keyword(s):  
Amino Acids ◽  
Common Ancestor ◽  
Gram Negative Bacteria ◽  
Citrobacter Freundii ◽  
Gene Encoding ◽  
Gram Negative ◽  
Putative Protein ◽  
Resistance Determinants ◽  
Flanking Sequences

A novel gene encoding a 6'-N-aminoglycoside acetyltransferase, aac(6')-In, has been cloned and sequenced from Citrobacter freundii 13996-19, a clinical isolate from Venezuela. This gene mediates resistance to amikacin, 2'-N-ethylnetilmicin, isepamicin, kanamycin, netilmicin, and tobramycin. The aac(6')-In gene is 573 nucleotides in length and encodes a putative protein of 190 amino acids. AAC(6')-In is most closely related to AAC(6')-Im and AAC(6')-Ie, demonstrating 64.4% and 62.3% similarity, respectively, at the protein level, suggesting these proteins share a common ancestor. The aac(6')-In flanking sequences demonstrated homology to integron- and transposon-related elements which are often found associated with resistance determinants. Hybridization studies performed with an intragenic probe specific for aac(6')-In indicate that this gene is prevalent within Venezuela but has not been observed outside of the country. Furthermore, the aac(6)-In gene was found in 10 different species of gram-negative bacteria.

scholarly journals Characterization of the 6'-N-aminoglycoside acetyltransferase gene aac(6')-Im [corrected] associated with a sulI-type integron.

1997 ◽  
Vol 41 (2) ◽  
pp. 314-318 ◽  
Author(s):  
E Hannecart-Pokorni ◽  
F Depuydt ◽  
L de wit ◽  
E van Bossuyt ◽  
J Content ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Resistance Gene ◽  
Citrobacter Freundii ◽  
Gram Negative ◽  
Gene Environment ◽  
Insert Region ◽  
Klebsiella Spp

The amikacin resistance gene aac(6')-Im [corrected] from Citrobacter freundii Cf155 encoding an aminoglycoside 6'-N-acetyltransferase was characterized. The gene was identified as a coding sequence of 521 bp located down-stream from the 5' conserved segment of an integron. The sequence of this aac(6')-Im [corrected] gene corresponded to a protein of 173 amino acids which possessed 64.2% identity in a 165-amino-acid overlap with the aac(6')-Ia gene product (F.C. Tenover, D. Filpula, K.L. Phillips, and J. J. Plorde, J. Bacteriol. 170:471-473, 1988). By using PCR, the aac(6')-Im [corrected] gene could be detected in 8 of 86 gram-negative clinical isolates from two Belgian hospitals, including isolates of Citrobacter, Klebsiella spp., and Escherichia coli. PCR mapping of the aac(6')-Im [corrected] gene environment in these isolates indicated that the gene was located within a sulI-type integron; the insert region is 1,700 bases long and includes two genes cassettes, the second being ant (3")-Ib.

Electrophysiological Characterization of Transport Across Outer Membrane Channels from Gram-Negative Bacteria in Their Native Environment

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Rémi Terrasse ◽  
Jayesh Arun Bafna ◽  
Lorraine Benier ◽  
Mathias Winterhalter
Keyword(s):  
Outer Membrane ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multi Drug Resistance ◽  
Gram Negative Bacteria ◽  
Membrane Channels ◽  
Gram Negative ◽  
Electrophysiological Characterization

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 <i>Escherichia coli</i>, 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 (<i>k<sub>on</sub></i>) and lower dissociation (<i>k<sub>off</sub></i>) 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.

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

2019 ◽  
Vol 6 (3) ◽  
pp. 467-478 ◽  
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

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

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. .

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

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.

scholarly journals Cloning, Nucleotide Sequence, and Expression of the Gene Encoding the Bacteriocin Boticin B from Clostridium botulinum Strain 213B

2000 ◽  
Vol 66 (12) ◽  
pp. 5480-5483 ◽  
Author(s):  
Sean S. Dineen ◽  
Marite Bradshaw ◽  
Eric A. Johnson
Keyword(s):  
Amino Acids ◽  
Nucleotide Sequence ◽  
Clostridium Botulinum ◽  
Shuttle Vector ◽  
Open Reading Frame ◽  
Gene Encoding ◽  
Reading Frame ◽  
Heat Stable ◽  
Group I

ABSTRACT Boticin B is a heat-stable bacteriocin produced byClostridium botulinum strain 213B that has inhibitory activity against various strains of C. botulinum and related clostridia. The gene encoding the bacteriocin was localized to a 3.0-kb HindIII fragment of an 18.8-kb plasmid, cloned, and sequenced. DNA sequencing revealed the boticin B structural gene,btcB, to be an open reading frame encoding 50 amino acids. A C. botulinum strain 62A transconjugant containing theHindIII fragment inserted into a clostridial shuttle vector expressed boticin B, although at much lower levels than those observed in C. botulinum 213B. To our knowledge, this is the first demonstration and characterization of a bacteriocin from toxigenic group I C. botulinum.

Val-Geninthiocin: Structure Elucidation and MSn Fragmentation of Thiopeptide Antibiotics Produced by Streptomyces sp. RSF18

2008 ◽  
Vol 63 (10) ◽  
pp. 1223-1230 ◽  
Author(s):  
Imran Sajid ◽  
Khaled A. Shaaban ◽  
Holm Frauendorf ◽  
Shahida Hasnain ◽  
Hartmut Laatscha
Keyword(s):  
Amino Acids ◽  
Biological Activity ◽  
Antifungal Activity ◽  
Structure Elucidation ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Unusual Amino Acids ◽  
Streptomyces Sp ◽  
Gram Positive Bacteria ◽  
Thiopeptide Antibiotics

AbstractVal-Geninthiocin (2), a new member of thiopeptide antibiotics, was isolated from the mycelium of Streptomyces sp. RSF18, along with the closely related geninthiocin (1) and the macrolide, chalcomycin. By intensive NMR and MS studies, Val-geninthiocin (2) was identified as desoxygeninthiocin, a thiopeptide, containing several oxazole and thiazole units and a number of unusual amino acids. Compound 2 shows potent activity against Gram-positive bacteria and minor antifungal activity, while it is not effective against Gram-negative bacteria or microalgae. Here we describe the fermentation, isolation and structure elucidation as well as the biological activity of 2.

scholarly journals Characterization of Certain Gram-Negative Bacteria from Surface Waters1

1965 ◽  
Vol 13 (4) ◽  
pp. 575-578 ◽  
Author(s):  
A. W. Hoadley ◽  
Elizabeth McCoy
Keyword(s):  
Gram Negative Bacteria ◽  
Gram Negative
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