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Search results for: pOET5 transfer plasmid

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#34566416   2021/09/18 To Up

IncN1 ST7 Epidemic Plasmid Carrying blaIMP-4 in One ST85-Type Clinical Isolate with Porin Deficiency.

is an opportunistic pathogen causing nosocomial infections. This study was designed to characterize the genomic features of a carbapenem-resistant strain and analyze its molecular characteristics.
Mingyue Sun, Weiqiang Xiao, Qingxia Xu

1558 related Products with: IncN1 ST7 Epidemic Plasmid Carrying blaIMP-4 in One ST85-Type Clinical Isolate with Porin Deficiency.

10 96 wells (1 kit)

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#34559608   2021/08/24 To Up

The Evolution of Plasmid Transfer Rate in Bacteria and Its Effect on Plasmid Persistence.

AbstractPlasmids are extrachromosomal segments of DNA that can transfer genes between bacterial cells. Many plasmid genes benefit bacteria but cause harm to human health by granting antibiotic resistance to pathogens. Transfer rate is a key parameter for predicting plasmid dynamics, but observed rates are highly variable, and the effects of selective forces on their evolution are unclear. We apply evolutionary analysis to plasmid conjugation models to investigate selective pressures affecting plasmid transfer rate, emphasizing host versus plasmid control, the costs of plasmid transfer, and the role of recipient cells. Our analyses show that plasmid-determined transfer rates can be predicted with three parameters (host growth rate, plasmid loss rate, and the cost of plasmid transfer on growth) under some conditions. We also show that low-frequency genetic variation in transfer rate can accumulate, facilitating rapid adaptation to changing conditions. Furthermore, reduced transfer rates due to host control have limited effects on plasmid prevalence until low enough to prevent plasmid persistence. These results provide a framework to predict plasmid transfer rate evolution in different environments and demonstrate the limited impact of host mechanisms to control the costs incurred when plasmids are present.
Richard J Sheppard, Timothy G Barraclough, Vincent A A Jansen

2473 related Products with: The Evolution of Plasmid Transfer Rate in Bacteria and Its Effect on Plasmid Persistence.

10 ug10 ug10 ug10 ug10 ug10 ug10 ug0.2ug/ul,20ul

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#34559044   // To Up

Flanker: a tool for comparative genomics of gene flanking regions.

Analysing the flanking sequences surrounding genes of interest is often highly relevant to understanding the role of mobile genetic elements (MGEs) in horizontal gene transfer, particular for antimicrobial-resistance genes. Here, we present Flanker, a Python package that performs alignment-free clustering of gene flanking sequences in a consistent format, allowing investigation of MGEs without prior knowledge of their structure. These clusters, known as 'flank patterns' (FPs), are based on Mash distances, allowing for easy comparison of similarity across sequences. Additionally, Flanker can be flexibly parameterized to fine-tune outputs by characterizing upstream and downstream regions separately, and investigating variable lengths of flanking sequence. We apply Flanker to two recent datasets describing plasmid-associated carriage of important carbapenemase genes ( and ) and show that it successfully identifies distinct clusters of FPs, including both known and previously uncharacterized structural variants. For example, Flanker identified four Tn profiles that could not be sufficiently characterized using TETyper or MobileElementFinder, demonstrating the utility of Flanker for flanking-gene characterization. Similarly, using a large (=226) European isolate dataset, we confirm findings from a previous smaller study demonstrating association between Tn and upregulation and demonstrate 17 FPs (compared to the 5 previously identified). More generally, the demonstration in this study that FPs are associated with geographical regions and antibiotic-susceptibility phenotypes suggests that they may be useful as epidemiological markers. Flanker is freely available under an MIT license at https://github.com/wtmatlock/flanker.
William Matlock, Samuel Lipworth, Bede Constantinides, Timothy E A Peto, A Sarah Walker, Derrick Crook, Susan Hopkins, Liam P Shaw, Nicole Stoesser

2658 related Products with: Flanker: a tool for comparative genomics of gene flanking regions.

50 ug25 µg50 ug1 module1 plate50 ug1 module50 ug50 ug50 ug50 ug

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#34557177   2021/09/07 To Up

Comparative Genomic Analysis Uncovered Evolution of Pathogenicity Factors, Horizontal Gene Transfer Events, and Heavy Metal Resistance Traits in Citrus Canker Bacterium subsp. .

Worldwide citrus production is severely threatened by Asiatic citrus canker which is caused by the proteobacterium subsp. . Foliar sprays of copper-based bactericides are frequently used to control plant bacterial diseases. Despite the sequencing of many strains, the genome diversity and distribution of genes responsible for metal resistance in subsp. strains from orchards with different management practices in Taiwan are not well understood. The genomes of three subsp. strains including one copper-resistant strain collected from farms with different management regimes in Taiwan were sequenced by Illumina and Nanopore sequencing and assembled into complete circular chromosomes and plasmids. CRISPR spoligotyping and phylogenomic analysis indicated that the three strains were located in the same phylogenetic lineages and shared ∼3,000 core-genes with published subsp. strains. These strains differed mainly in the CRISPR repeats and pathogenicity-related plasmid-borne transcription activator-like effector (TALE)-encoding genes. The copper-resistant strain has a unique, large copper resistance plasmid due to an unusual ∼40 kbp inverted repeat. Each repeat contains a complete set of the gene cluster responsible for copper and heavy metal resistance. Conversely, the copper sensitive strains carry no metal resistance genes in the plasmid. Through comparative analysis, the origin and evolution of the metal resistance clusters was resolved. Chromosomes remained constant among three strains collected in Taiwan, but plasmids likely played an important role in maintaining pathogenicity and developing bacterial fitness in the field. The evolution of pathogenicity factors and horizontal gene transfer events were observed in the three strains. These data suggest that agricultural management practices could be a potential trigger for the evolution of citrus canker pathogens. The decrease in the number of CRISPR repeats and genes might be the result of adaptation to a less stressful environment. The metal resistance genes in the copper resistant strain likely originated from the Mauritian strain not the local copper-resistant strain. This study highlights the importance of plasmids as 'vehicles' for exchanging genetic elements between plant pathogenic bacteria and contributing to bacterial adaptation to the environment.
Chien-Jui Huang, Ting-Li Wu, Po-Xing Zheng, Jheng-Yang Ou, Hui-Fang Ni, Yao-Cheng Lin

1379 related Products with: Comparative Genomic Analysis Uncovered Evolution of Pathogenicity Factors, Horizontal Gene Transfer Events, and Heavy Metal Resistance Traits in Citrus Canker Bacterium subsp. .

300 units5ug96T100.00 ug96T

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#34552574   2021/09/06 To Up

Genetic Analysis of Carrying Plasmids From Gram-Negative Bacteria in a Dutch Tertiary Care Hospital: Evidence for Intrapatient and Interspecies Transmission Events.

The role of plasmids in the complex pandemic of antimicrobial resistance is increasingly being recognized. In this respect, multiple mobile colistin resistance () gene-carrying plasmids have been described. However, the characteristics and epidemiology of these plasmids within local healthcare settings are largely unknown. We retrospectively characterized the genetic composition and epidemiology of plasmids from positive bacterial isolates identified from patients from a large academic hospital in the Netherlands. Clinical Gram-negative bacteria with an MIC > 2 μg/mL for colistin, obtained from patients hospitalized at the Erasmus MC University Medical Center Rotterdam during the years 2010-2018, were screened for presence of the gene. Extracted plasmids from -positive isolates were sequenced using a combination of short- and long-read sequencing platforms, characterized by incompatibility type and genetic composition and compared to publicly available carrying plasmid sequences. In 21 isolates from 14 patients, was located on a plasmid. These plasmids were of diverse genetic background involving Inc types IncX4, IncI2(delta), IncHI2, as well as double Inc types IncHI2/IncN and IncHI2/IncQ. -carrying plasmids were found in , and within the chromosome of an ST147 isolate. In depth analysis indicated intrapatient, interpatient, and interspecies transmission events of carrying plasmids. In addition, our results show that the gene resides in a rich environment full of other ( negative) plasmids and of many different Inc types, enabling interplasmidal transfer events and facilitating widespread dissemination of the gene. Multiple -carrying plasmid transmission events had likely occurred among isolates from hospitalized patients. Recognition and identification of plasmid transmission events within hospitals is necessary in order to design and implement effective infection control measures.
Nikolaos Strepis, Anne F Voor In 't Holt, Margreet C Vos, Willemien H A Zandijk, Astrid P Heikema, John P Hays, Juliëtte A Severin, Corné H W Klaassen

1373 related Products with: Genetic Analysis of Carrying Plasmids From Gram-Negative Bacteria in a Dutch Tertiary Care Hospital: Evidence for Intrapatient and Interspecies Transmission Events.

1 mL1 mg0.25 ml1 mL1 mg0.25 ml1 mg0.2 mg1 KIT

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#34547258   2021/09/20 To Up

Dynamic mRNA polyplexes benefit from bioreducible cleavage sites for in vitro and in vivo transfer.

Currently, messenger RNA (mRNA)-based lipid nanoparticle formulations revolutionize the clinical field. Cationic polymer-based complexes (polyplexes) represent an alternative compound class for mRNA delivery. After establishing branched polyethylenimine with a succinylation degree of 10% (succPEI) as highly effective positive mRNA transfection standard, a diverse library of PEI-like peptides termed sequence-defined oligoaminoamides (OAAs) was screened for mRNA delivery. Notably, sequences, which had previously been identified as potent plasmid DNA (pDNA) or small-interfering RNA (siRNA) carriers, displayed only moderate mRNA transfection activity. A second round of screening combined the cationizable building block succinoyl tetraethylene pentamine and histidines for endosomal buffering, tyrosine tripeptides and various fatty acids for mRNA polyplex stabilization, as well as redox-sensitive units for programmed intracellular release. For the tested OAA carriers, balancing of extracellular stability, endosomal lytic activity, and intracellular release capability was found to be of utmost importance for optimum mRNA transfection efficiency. OAAs with T-shape topology containing two oleic acids as well-stabilizing fatty acids, attached via a dynamic bioreducible building block, displayed superior activity with up to 1000-fold increased transfection efficiency compared to their non-reducible analogs. In the absence of the dynamic linkage, incorporation of shorter less stabilizing fatty acids could only partly compensate for mRNA delivery. Highest GFP expression and the largest fraction of transfected cells (96%) could be detected for the bioreducible OAA with incorporated histidines and a dioleoyl motif, outperforming all other tested carriers as well as the positive control succPEI. The good in vitro performance of the dynamic lead structure was verified in vivo upon intratracheal administration of mRNA complexes in mice.
Ana Krhač Levačić, Simone Berger, Judith Müller, Andrea Wegner, Ulrich Lächelt, Christian Dohmen, Carsten Rudolph, Ernst Wagner

1569 related Products with: Dynamic mRNA polyplexes benefit from bioreducible cleavage sites for in vitro and in vivo transfer.

500 MG96 tests48 samples 100 G50 ug1mg 1 G10mg

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#34536683   2021/09/04 To Up

Inhibited conjugative transfer of antibiotic resistance genes in antibiotic resistant bacteria by surface plasma.

Antibiotic resistant bacteria (ARB) and resistance genes (ARGs) are emerging environmental pollutants with strong pathogenicity. In this study, surface plasma was developed to inactivate the donor ARB with Escherichia coli (AR E. coli) as a model, eliminate ARGs, and inhibit conjugative transfer of ARGs in water, highlighting the influences of concomitant inorganic ions. Surface plasma oxidation significantly inactivated AR E. coli, eliminated ARGs, and inhibited conjugative transfer of ARGs, and the presence of NO, Cu, and Fe all promoted these processes, and SO did not have distinct effect. Approximately 4.5log AR E. coli was inactivated within 10 min treatment, and it increased to 7.4log AR E. coli after adding Fe. Integrons intI1 decreased by 3.10log (without Fe) and 4.43log (adding Fe); the addition of Fe in the surface plasma induced 99.8% decline in the conjugative transfer frequency. The inhibition effects on the conjugative transfer of ARGs were mainly attributed to the reduced reactive oxygen species levels, decreased DNA damage-induced response, decreased intercellular contact, and down-regulated expression of plasmid transfer genes. This study disclosed underlying mechanisms for inhibiting ARGs transfer, and supplied a prospective technique for ARGs control.
Hu Li, Ruiying Song, Yangyang Wang, Rongwei Zhong, Ying Zhang, Jian Zhou, Tiecheng Wang, Hanzhong Jia, Lingyan Zhu

1508 related Products with: Inhibited conjugative transfer of antibiotic resistance genes in antibiotic resistant bacteria by surface plasma.



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#34533680   2021/09/17 To Up

Generation of stable cell lines using readthrough expression from lentiviral integration.

Lentiviral infection is often used to integrate genetic material into cells to stably express transgenes of interest. Depending on the location of integration into the host genome, readthrough expression of the lentiviral cargo can occur via an upstream endogenous promoter, which is typically an unwanted phenomenon because it can result in dysfunctional expression. The purpose of this study was to demonstrate that readthrough expression can be a wanted phenomenon for expressing functional proteins while at the same time reducing the size of the lentiviral transfer plasmid. Readthrough expression was used to generate HEK293 cell lines stably expressing fluorescent reporter proteins, reporter protein-antibiotic resistance fusion proteins for selection, and the vascular endothelial growth factor receptor 2. The generated proteins were all functional, as demonstrated by their ability to fluoresce, confer antibiotic resistance, and participate in receptor-mediated signalling, respectively. Therefore, we suggest that the mechanism of readthrough expression may have further applications in the expression of larger genes or genetic circuits (e.g. cell-based therapeutics), where the lentiviral cargo limit is stretched to the maximum.
Anish Jadav, Kevin Truong

2565 related Products with: Generation of stable cell lines using readthrough expression from lentiviral integration.

One Vial: 5 X 10^6 Cells10 plates100 platesvial100 plates10 ug1.5x10(6) cells

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#34531844   2021/08/31 To Up

Identification and Characterization of a Novel Aminoglycoside 3''-Nucleotidyltransferase, ANT(3'')-IId, From .

A novel plasmid-encoded aminoglycoside 3''-nucleotidyltransferase ANT(3")-IId, was discovered in strain H7 isolated from a chick on an animal farm in Wenzhou, China. The whole-genome of H7 consisted of one chromosome and five plasmids (pH7-250, pH7-108, pH7-68, pH7-48, and pH7-11). was identified as being encoded on pH7-250, sharing the highest amino acid identity of 50.64% with a function-known resistance gene, (KB849358.1). Susceptibility testing and enzyme kinetic parameter analysis were conducted to determine the function of the aminoglycoside 3"-nucleotidyltransferase. The gene conferred resistance to spectinomycin and streptomycin [the minimum inhibitory concentration (MIC) levels of both increased 16-fold compared with the control strain]. Consistent with the MIC data, kinetic analysis revealed a narrow substrate profile including spectinomycin and streptomycin, with / ratios of 4.99 and 4.45×10M S, respectively. Sequencing analysis revealed that the gene was associated with insertion sequences (IS) element [ΔIS-ΔIS-hp-orf-orf-orf1-], and were identified in plasmids from various species. This study of the novel aminoglycoside 3"-nucleotidyltranferase ANT(3")-IId helps us further understand the functional and sequence characteristics of aminoglycoside 3"-nucleotidyltranferases, highlights the risk of resistance gene transfer among species and suggests that attention should be given to the emergence of new aminoglycoside 3"-nucleotidyltranferase genes.
Jialei Liang, Kexin Zhou, Qiaoling Li, Xu Dong, Peiyao Zhang, Hongmao Liu, Hailong Lin, Xueya Zhang, Junwan Lu, Xi Lin, Kewei Li, Teng Xu, Hailin Zhang, Qiyu Bao, Mei Zhu, Yunliang Hu, Ping Ren

2824 related Products with: Identification and Characterization of a Novel Aminoglycoside 3''-Nucleotidyltransferase, ANT(3'')-IId, From .

25 mg50 ug 100 mg0.1ml (1mg/ml) 25 MG100ug10 mg100ug25 mg1000 tests100ul

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#34530277   2021/09/09 To Up

Biochar effectively inhibits the horizontal transfer of antibiotic resistance genes via transformation.

The rapid spread of antibiotic resistance genes (ARGs) has posed a risk to human health. Here, the effects of biochar (BC) on the horizontal transfer of ARG-carrying plasmids to Escherichia coli via transformation were systematically investigated. BC could significantly inhibit the transformation of ARGs and the inhibition degree increased with pyrolysis temperature. Rice straw-derived BC showed a stronger inhibitory effect on the transformation of ARGs than that of peanut shell-derived BC from the same pyrolysis temperature. The inhibitory effect of BC from low pyrolysis temperature (300 ℃) was mainly caused by BC dissolutions, while it was mainly attributed to BC solids for high pyrolysis temperature (700 ℃) BC. BC dissolutions could induce intramolecular condensation and even agglomeration of plasmids, hindering their transformation into competent bacteria. The cell membrane permeability was slightly decreased in BC dissolutions, which might also contribute to the inhibitory effect. Plasmid can be adsorbed by BC solids and the adsorption increased with BC pyrolysis temperature. Meanwhile, BC-adsorbed plasmid could hardly be transformed into E. coli. BC solids could also deactivate E. coli and thereby inhibit their uptake of ARGs. These findings provide a way using BC to limit the spread of ARGs in the environment.
Jing Fang, Liang Jin, Qingkang Meng, Shengdao Shan, Dengjun Wang, Daohui Lin

2407 related Products with: Biochar effectively inhibits the horizontal transfer of antibiotic resistance genes via transformation.

100 ug/vial0.2 mg1 x 10^6 cells/vial100ug Lyophilized100 ug/vial 25UG1100μg/vial1 g100 ug/vial

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