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Plasmid curing

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https://www.readbyqxmd.com/read/28433723/elimination-of-the-cryptic-plasmid-in-leuconostoc-citreum-by-crispr-cas9-system
#1
Ye-Ji Jang, Seung-Oh Seo, Seul-Ah Kim, Ling Li, Tae-Jip Kim, Sun Chang Kim, Yong-Su Jin, Nam Soo Han
Leuconostoc spp. are important lactic acid bacteria for the fermentation of foods. In particular, L. citreum strains isolated from various foods have been used as host strains for genetic and metabolic engineering studies. In order to develop a food-grade genetic engineering system, L. citreum CB2567 was isolated from Kimchi. However, the isolated bacterium contained a cryptic plasmid which was difficult to eliminate. As the existence of the plasmid might hinder strain engineering, we eliminated the plasmid using an RNA-guided DNA endonuclease CRISPR/Cas9 system...
April 19, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28432094/the-use-of-synthetic-hybrid-strains-to-determine-the-role-of-replicon-3-in-virulence-of-the-burkholderia-cepacia-complex
#2
Kirsty Agnoli, Roman Freitag, Margarida C Gomes, Christian Jenul, Angela Suppiger, Olga Mannweiler, Carmen Frauenknecht, Daniel Janser, Annette C Vergunst, Leo Eberl
The Burkholderia cepacia complex (Bcc) displays a wealth of metabolic diversity with great biotechnological potential, but the utilisation of these bacteria is limited by their opportunistic pathogenicity to humans. The third replicon of the Bcc, megaplasmid pC3 (0.5 to 1.4 Mb, previously chromosome 3), is important for various phenotypes, including virulence, antifungal and proteolytic activities, and the utilization of certain substrates. Approximately half of plasmid pC3 is well-conserved throughout sequenced Bcc members, while the other half is not...
April 21, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28330200/use-of-ca-alginate-immobilized-pseudomonas-aeruginosa-for-repeated-batch-and-continuous-degradation-of-endosulfan
#3
Vijayalakshmi Pradeep, Usha Malavalli Subbaiah
The current investigation is taken up with the aim of studying repeated batch and continuous degradation of Endosulfan, using Ca-alginate immobilized cells of Pseudomonas aeruginosa isolated from an agricultural soil. The work involves the study of genes and enzymes involved in the degradation of the pesticide and was carried out with an objective of reducing the toxicity of Endosulfan by degrading it to less toxic metabolites. The long-term stability of Endosulfan degradation was studied during its repeated batch degradation, carried out over a period of 35 days...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330180/biodegradation-of-propiconazole-by-newly-isolated-burkholderia-sp-strain-bbk_9
#4
Praveen Satapute, Basappa Kaliwal
The isolation of propiconazole (PCZ) degrading bacterium BBK_9 strain was done from paddy soil, and it was identified as Burkholderia sp. based on the morphological characteristics and biochemical properties combined with 16S rRNA gene sequencing analysis. It has been seen that the factors such as temperature and pH influence the biodegradation process. The role of plasmid was studied in the degradation process by plasmid curing method. The PCZ acts as the sole carbon source and as energy substrate which can be utilized by the strain for its growth in Mineral salt medium and degraded 8...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28279738/modifying-plasmid-loaded-hsa-nanoparticles-with-cell-penetrating-peptides-cellular-uptake-and-enhanced-gene-delivery
#5
J Mesken, A Iltzsche, D Mulac, K Langer
Gene therapy bears great potential for the cure of a multitude of human diseases. Research efforts focussed on the use of viral delivery vectors in the past decades, neglecting non-viral gene therapies of physical or chemical origin due to low transfection efficiency. However, side effects such as activation of oncogenes and inflammatory reactions upon immune cell activation are major obstacles impeding the clinical applicability of viral gene therapy vectors. The aim of this study was the development of a non-viral gene delivery system based on plasmid-loaded human serum albumin nanoparticles, which are biocompatible, biodegradable, and non-toxic in relevant concentrations...
March 6, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/28277944/the-galleria-mellonella-larvae-as-an-in-vivo-model-for-evaluation-of-shigella-virulence
#6
Shoshana Barnoy, Hanan Gancz, Yuewei Zhu, Cary L Honnold, Daniel V Zurawski, Malabi M Venkatesan
Shigella spp. causing bacterial diarrhea and dysentery are human enteroinvasive bacterial pathogens that are orally transmitted through contaminated food and water and cause bacillary dysentery. Although natural Shigella infections are restricted to humans and primates, several smaller animal models are used to analyze individual steps in pathogenesis. No animal model fully duplicates the human response and sustaining the models requires expensive animals, costly maintenance of animal facilities, veterinary services and approved animal protocols...
February 13, 2017: Gut Microbes
https://www.readbyqxmd.com/read/28261566/efflux-pump-overexpression-contributes-to-tigecycline-heteroresistance-in-salmonella-enterica-serovar-typhimurium
#7
Yi Chen, Daxing Hu, Qijing Zhang, Xiao-Ping Liao, Ya-Hong Liu, Jian Sun
Bacterial heteroresistance has been identified in several combinations of bacteria and antibiotics, and it complicated the therapeutic strategies. Tigecycline is being used as one of the optimal options for the treatment of infections caused by multidrug-resistant Salmonella. This study investigated whether heterorresistance to tigecycline exists in a Salmonella enterica serovar Typhimurium strain harboring the oqxAB-bearing IncHI2 plasmid pHXY0908. MIC and population analyses were performed to evaluate population-wide susceptibility to tigecycline...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28245276/plasmid-interference-for-curing-antibiotic-resistance-plasmids-in-vivo
#8
Muhammad Kamruzzaman, Shereen Shoma, Christopher M Thomas, Sally R Partridge, Jonathan R Iredell
Antibiotic resistance increases the likelihood of death from infection by common pathogens such as Escherichia coli and Klebsiella pneumoniae in developed and developing countries alike. Most important modern antibiotic resistance genes spread between such species on self-transmissible (conjugative) plasmids. These plasmids are traditionally grouped on the basis of replicon incompatibility (Inc), which prevents coexistence of related plasmids in the same cell. These plasmids also use post-segregational killing ('addiction') systems, which poison any bacterial cells that lose the addictive plasmid, to guarantee their own survival...
2017: PloS One
https://www.readbyqxmd.com/read/28238083/elimination-of-indigenous-linear-plasmids-in-streptomyces-hygroscopicus-var-jinggangensis-and-streptomyces-sp-fr008-to-increase-validamycin-a-and-candicidin-productivities
#9
Chenyang Lu, Hang Wu, Xiurong Su, Linquan Bai
Giant linear plasmids, which replicate independently of the chromosomes, widely exist in actinobacteria. Previous studies mostly focused on the replication and evolution of the linear plasmids or the secondary metabolite gene clusters and the resistance gene clusters therein. However, the relationships of the linear plasmids to the productivities of secondary metabolites have not been studied. In this work, we developed a method to eliminate the indigenous linear plasmid pSHJG1 in Streptomyces hygroscopicus var...
February 25, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28192223/emergence-of-multi-drug-resistance-among-soil-bacteria-exposing-to-insecticides
#10
Kirubakaran Rangasamy, Murugan Athiappan, Natarajan Devarajan, Javid A Parray
Impacts of pesticide exposure on the soil microbial flora and cross resistance to antibiotics have not been well documented. Development of antibiotic resistance is a common issue among soil bacteria which are exposing to pesticides continuously at sub-lethal concentration. The present study was focused to evaluate the correlation between pesticide exposures and evolution of multi drug resistance among isolates collected from soil applied with insecticides. Twenty five insecticide (Monochrotophos) degrading bacteria were isolated from contaminated agricultural soil...
February 10, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28179404/a-plasmodium-vivax-plasmid-dna-and-adenovirus-vectored-malaria-vaccine-encoding-blood-stage-antigens-ama1-and-msp142-in-a-prime-boost-heterologous-immunization-regimen-partially-protects-aotus-monkeys-against-blood-stage-challenge
#11
Nicanor Obaldia, Michael G Stockelman, William Otero, Jennifer A Cockrill, Harini Ganeshan, Esteban N Abot, Jianfeng Zhang, Keith Limbach, Yupin Charoenvit, Denise L Doolan, De-Chu C Tang, Thomas L Richie
Malaria is caused by parasites of the genus Plasmodium, which are transmitted to humans by the bites of Anopheles mosquitoes. After the elimination of Plasmodium falciparum, it is predicted that Plasmodium vivax will remain an important cause of morbidity and mortality outside Africa, stressing the importance of developing a vaccine against P. vivax malaria. In this study, we assessed the immunogenicity and protective efficacy of two P. vivax antigens, apical membrane antigen 1 (AMA1) and the 42-kDa C-terminal fragment of merozoite surface protein 1 (MSP142) in a plasmid recombinant DNA prime/adenoviral (Ad) vector boost regimen in Aotus monkeys...
April 2017: Clinical and Vaccine Immunology: CVI
https://www.readbyqxmd.com/read/28161562/hindrance-of-1-4-dioxane-biodegradation-in-microcosms-biostimulated-with-inducing-or-non-inducing-auxiliary-substrates
#12
Mengyan Li, Yuanyuan Liu, Ya He, Jacques Mathieu, James Hatton, William DiGuiseppi, Pedro J J Alvarez
A microcosm study was conducted to assess two biostimulation strategies (relative to natural attenuation) to bioremediate 1,4-dioxane contamination at a site in west Texas. Dioxane concentrations were relatively low (<300 μg/L), which represents a potential challenge to sustain and induce specific degraders. Thus, biostimulation was attempted with an auxiliary substrate known to induce dioxane-degrading monooxygenases (i.e., tetrahydrohyran [THF]) or with a non-inducing growth substrate (1-butanol [1-BuOH])...
January 26, 2017: Water Research
https://www.readbyqxmd.com/read/28154555/key-impact-of-an-uncommon-plasmid-on-bacillus-amyloliquefaciens-subsp-plantarum-s499-developmental-traits-and-lipopeptide-production
#13
Giulia Molinatto, Laurent Franzil, Sébastien Steels, Gerardo Puopolo, Ilaria Pertot, Marc Ongena
The rhizobacterium Bacillus amyloliquefaciens subsp. plantarum S499 (S499) is particularly efficient in terms of the production of cyclic lipopeptides, which are responsible for the high level of plant disease protection provided by this strain. Sequencing of the S499 genome has highlighted genetic differences and similarities with the closely related rhizobacterium B. amyloliquefaciens subsp. plantarum FZB42 (FZB42). More specifically, a rare 8008 bp plasmid (pS499) harboring a rap-phr cassette constitutes a major distinctive element between S499 and FZB42...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28108611/reversal-of-diabetes-in-nod-mice-by-clinical-grade-proinsulin-and-il-10-secreting-lactococcus-lactis-in-combination-with-low-dose-anti-cd3-depends-on-the-induction-of-foxp3-positive-t-cells
#14
Tatiana Takiishi, Dana Paulina Cook, Hannelie Korf, Guido Sebastiani, Francesca Mancarella, João Paulo Monteiro Carvalho Mori Cunha, Clive Wasserfall, Noelia Casares, Juan José Lasarte, Lothar Steidler, Pieter Rottiers, Francesco Dotta, Conny Gysemans, Chantal Mathieu
The introduction of β-cell autoantigens via the gut through Lactococcus lactis (L. lactis) has been demonstrated to be a promising approach for diabetes reversal in NOD mice. Here we show that a combination therapy of low-dose anti-CD3 with a clinical-grade self-containing L. lactis, appropriate for human application, secreting human proinsulin and interleukin-10, cured 66% of mice with new-onset diabetes, which is comparable to therapy results with plasmid-driven L. lactis Initial blood glucose concentrations (<350 mg/dL) and insulin autoantibody positivity were predictors of the stable reversal of hyperglycemia, and decline in insulin autoantibody positivity was an immune biomarker of therapeutic outcome...
February 2017: Diabetes
https://www.readbyqxmd.com/read/27941999/plasmid-complement-of-lactococcus-lactis-ncdo712-reveals-a-novel-pilus-gene-cluster
#15
Mariya Tarazanova, Marke Beerthuyzen, Roland Siezen, Marcela M Fernandez-Gutierrez, Anne de Jong, Sjoerd van der Meulen, Jan Kok, Herwig Bachmann
Lactococcus lactis MG1363 is an important gram-positive model organism. It is a plasmid-free and phage-cured derivative of strain NCDO712. Plasmid-cured strains facilitate studies on molecular biological aspects, but many properties which make L. lactis an important organism in the dairy industry are plasmid encoded. We sequenced the total DNA of strain NCDO712 and, contrary to earlier reports, revealed that the strain carries 6 rather than 5 plasmids. A new 50-kb plasmid, designated pNZ712, encodes functional nisin immunity (nisCIP) and copper resistance (lcoRSABC)...
2016: PloS One
https://www.readbyqxmd.com/read/27902432/self-replicating-shuttle-vectors-based-on-pans-a-small-endogenous-plasmid-of-the-unicellular-cyanobacterium-synechococcus-elongatus-pcc-7942
#16
You Chen, Arnaud Taton, Michaela Go, Ross E London, Lindsey M Pieper, Susan S Golden, James W Golden
To facilitate development of synthetic biology tools for genetic engineering of cyanobacterial strains, we constructed pANS-derived self-replicating shuttle vectors that are based on the minimal replication element of the Synechococcus elongatus strain PCC 7942 plasmid pANS. To remove the possibility of homologous recombination events between the shuttle plasmids and the native pANS plasmid, the endogenous pANS was cured through plasmid incompatibility-mediated spontaneous loss. A heterologous toxin-antitoxin cassette was incorporated into the shuttle vectors for stable plasmid maintenance in the absence of antibiotic selection...
December 2016: Microbiology
https://www.readbyqxmd.com/read/27900888/using-the-crispr-cas9-system-to-eliminate-native-plasmids-of-zymomonas-mobilis-zm4
#17
Qing-Hua Cao, Huan-Huan Shao, Hui Qiu, Tao Li, Yi-Zheng Zhang, Xue-Mei Tan
The CRISPR/Cas system can be used to simply and efficiently edit the genomes of various species, including animals, plants, and microbes. Zymomonas mobilis ZM4 is a highly efficient, ethanol-producing bacterium that contains five native plasmids. Here, we constructed the pSUZM2a-Cas9 plasmid and a single-guide RNA expression plasmid. The pSUZM2a-Cas9 plasmid was used to express the Cas9 gene cloned from Streptococcus pyogenes CICC 10464. The single-guide RNA expression plasmid pUC-T7sgRNA, with a T7 promoter, can be used for the in vitro synthesis of single-guide RNAs...
March 2017: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/27882060/simple-and-reliable-method-to-quantify-the-hepatitis-b-viral-load-and-replicative-capacity-in-liver-tissue-and-blood-leukocytes
#18
Claudia Minosse, Sabrina Coen, Ubaldo Visco Comandini, Raffaella Lionetti, Marzia Montalbano, Stefano Cerilli, Donatella Vincenti, Andrea Baiocchini, Maria R Capobianchi, Stefano Menzo
BACKGROUND: A functional cure of chronic hepatitis B (CHB) is feasible, but a clear view of the intrahepatic viral dynamics in each patient is needed. Intrahepatic covalently closed circular DNA (cccDNA) is the stable form of the viral genome in infected cells, and represents the ideal marker of parenchymal colonization. Its relationships with easily accessible peripheral parameters need to be elucidated in order to avoid invasive procedures in patients. OBJECTIVES: The goal of this study was to design, set up, and validate a reliable and straightforward method for the quantification of the cccDNA and total DNA of the hepatitis B virus (HBV) in a variety of clinical samples...
October 2016: Hepatitis Monthly
https://www.readbyqxmd.com/read/27786189/crispr-rna-guided-foki-nucleases-repair-a-pah-variant-in-a-phenylketonuria-model
#19
Yi Pan, Nan Shen, Sabine Jung-Klawitter, Christian Betzen, Georg F Hoffmann, Jörg D Hoheisel, Nenad Blau
The CRISPR/Cas9 system is a recently developed genome editing technique. In this study, we used a modified CRISPR system, which employs the fusion of inactive Cas9 (dCas9) and the FokI endonuclease (FokI-dCas9) to correct the most common variant (allele frequency 21.4%) in the phenylalanine hydroxylase (PAH) gene - c.1222C>T (p.Arg408Trp) - as an approach toward curing phenylketonuria (PKU). PKU is the most common inherited diseases in amino acid metabolism. It leads to severe neurological and neuropsychological symptoms if untreated or late diagnosed...
October 27, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27746775/inchi2-plasmids-are-predominant-in-antibiotic-resistant-salmonella-isolates
#20
Wenyao Chen, Tingzi Fang, Xiujuan Zhou, Daofeng Zhang, Xianming Shi, Chunlei Shi
The wide usage of antibiotics contributes to the increase in the prevalence of antibiotic-resistant Salmonella. Plasmids play a critical role in horizontal transfer of antibiotic resistance markers in Salmonella. This study aimed to screen and characterize plasmid profiles responsible for antibiotic resistance in Salmonella and ultimately to clarify the molecular mechanism of transferable plasmid-mediated antibiotic resistance. A total of 226 Salmonella isolates were examined for antimicrobial susceptibility by a disk diffusion method...
2016: Frontiers in Microbiology
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