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https://www.readbyqxmd.com/read/28922869/genome-organization-of-sphingobium-indicum-b90a-an-archetypal-hexachlorocyclohexane-hch-degrading-genotype
#1
Helianthous Verma, Abhay Bajaj, Roshan Kumar, Jasvinder Kaur, Shailly Anand, Namita Nayyar, Akshita Puri, Yogendra Singh, Jitendra P Khurana, Rup Lal
Among sphingomonads, Sphingobium indicum B90A is widely investigated for its ability to degrade a manmade pesticide, γ-hexachlorocyclohexane (γ-HCH) and its isomers (α-, β-, δ-, and ε-HCH). In this study, complete genome of strain B90A was constructed using Single Molecule Real Time Sequencing (SMRT) and Illumina platform. The complete genome revealed that strain B90A harbors four replicons: one chromosome (3,654,322 bp) and three plasmids designated as pSRL1 (139,218 bp), pSRL2 (108,430 bp) and pSRL3 (43,761 bp)...
September 1, 2017: Genome Biology and Evolution
https://www.readbyqxmd.com/read/28922058/evolution-of-regions-containing-antibiotic-resistance-genes-in-fii-2-fib-1-colv-colla-virulence-plasmids
#2
Robert A Moran, Ruth M Hall
Three ColV virulence plasmids carrying antibiotic resistance genes were assembled from draft genome sequences of commensal ST95, ST131, and ST2705 Escherichia coli isolates from healthy Australians. Plasmids pCERC4, pCERC5, and pCERC9 include almost identical backbones containing FII-2 and FIB-1 replicons and the conserved ColV virulence region with an additional ColIa determinant. Only pCERC5 includes a complete, uninterrupted F-like transfer region and was able to conjugate. pCERC5 and pCERC9 contain Tn1721, carrying the tet(A) tetracycline resistance determinant in the same location, with Tn2 (blaTEM; ampicillin resistance) interrupting the Tn1721 in pCERC5...
September 18, 2017: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
https://www.readbyqxmd.com/read/28922026/characterization-of-quinolone-resistance-in-salmonella-enterica-from-farm-animals-in-china
#3
Ting-Ting Cao, Guo-Hui Deng, Liang-Xing Fang, Run-Shi Yang, Jian Sun, Ya-Hong Liu, Xiao-Ping Liao
This study was focused on the prevalence and antimicrobial susceptibilities of Salmonella directly isolated at animal clinics in Guangdong, People's Republic of China. The isolation rates from chickens, ducks, and pigs were 11.3% (11 of 97 samples), 15.4% (53 of 344 samples), and 3.0% (13 of 434 samples), respectively. Among the 77 Salmonella enterica isolates, the most predominant serovar was Typhimurium (81.8%, 63 isolates), followed by serovars Meleagridis (2.6%, 2 isolates) and Abaetetuba (1.3%, 1 isolate)...
September 18, 2017: Journal of Food Protection
https://www.readbyqxmd.com/read/28921745/transcription-factor-ccaat-enhancer-binding-protein-%C3%AE-upregulates-microrna-let-7f-1-in-human-endocervical-cells
#4
Kanchana Ayyar, Kudumula Venkata Rami Reddy
PROBLEM: In endocervical epithelial cells (End1/E6E7), miRNA let-7f plays an important role in the control of innate immunity. The underlying molecular mechanism for let-7f regulation in these cells remains largely unclear. METHODS OF STUDY: let-7f was knocked down in End1/E6E7 cells using siRNA, and differential gene expression was analyzed by microarray. Differentially expressed genes were validated by qPCR and Western blot. Expression of let-7f was studied by qPCR after inhibition of C/EBPβ with betulinic acid (BA) and pCMVβ plasmid and after overexpression of C/EBPβ with pCMVβ+ plasmid...
September 16, 2017: American Journal of Reproductive Immunology: AJRI
https://www.readbyqxmd.com/read/28921433/large-scale-transient-transfection-of-suspension-mammalian-cells-for-vlp-production
#5
Laura Cervera, Amine A Kamen
Large-scale transient transfection of mammalian cell suspension cultures enables the production of biological products in sufficient quantity and under stringent quality attributes to perform accelerated in vitro evaluations and has the potential to support preclinical or even clinical studies. Here we describe the methodology to produce VLPs in a 3L bioreactor, using suspension HEK 293 cells and PEIPro as a transfection reagent. Cells are grown in the bioreactor to 1 × 10(6) cells/mL and transfected with a plasmid DNA-PEI complex at a ratio of 1:2...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28920846/detection-of-the-incx3-plasmid-carrying-blakpc-3-in-a-serratia-marcescens-strain-isolated-from-a-kidney-liver-transplanted-patient
#6
Floriana Gona, Carla Caio, Gioacchino Iannolo, Francesco Monaco, Giuseppina Di Mento, Nicola Cuscino, Ignazio Fontana, Giovanna Panarello, Gaetano Maugeri, Maria Lina Mezzatesta, Stefania Stefani, Pier Giulio Conaldi
Dissemination of resistance to carbapenems among Enterobacteriaceae through plasmids is an increasingly important concern in health care worldwide. Here we report the first description of an IncX3 plasmid carrying the blaKPC-3 gene in a strain of Serratia marcescens isolated from a kidney-liver transplanted patient at the transplantation centre ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo, Italy). To localize the transposable element containing the resistance-associated gene Next-Generation Sequencing of the bacterial DNA was performed...
September 18, 2017: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/28920402/-preparation-and-transformation-optimization-for-supercompetent-b-subtilis-sck6-cells
#7
Xinzhi Li, Zhenghui Lu, Yuling Zhou, Shiyu Li, Guimin Zhang
Bacillus subtilis is Gram-positive aerobic bacterium and widely used as a heterologous protein expression host because of its safety and high protein secretion property. However, comparing to Escherichia coli, the low transformation efficiency limits the application of B. subtilis as a host cell for directed evolution of heterologous enzymes. Therefore, we optimized the competent cell preparation conditions for conventional plasmid, including the alteration of the medium, the concentration of inducer, the plasmid type, and other parameters...
April 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28920397/-effects-of-interleukin-34-expressed-by-human-bone-marrow-derived-mesenchymal-stem-cells-on-thp-1-cells
#8
Xueming Gao, Shun Zhang, Zhaojun Xu, Yan Shan, Zihong Wang, Ting Cai, Xue Jiang
To construct recombinant eukaryotic expression plasmid vector of human IL-34 gene, and to study the effects of IL-34 expressed by human bone marrow-derived mesenchymal stem cells (hBM-MSCs) on THP-1 cells. Full-length IL-34 encoding sequence was amplified by PCR. And this fragment was cloned into the plasmid pIRES2-EGFP. Western blotting and ELISA were used to analyze the expression of IL-34 in hBM-MSCs. THP-1 cells were cultured with hBM-MSCs medium containing IL-34 protein. Real-time PCR detected the effects of IL-34 on the expression of IL-10 and TNFα in THP-1 cells...
April 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28920389/-research-progress-in-multi-enzyme-regulation-of-genetically-engineered-bacteria-producing-lycopene
#9
Jiali Xu, Siyi Zuo, Chengjia Xie, Ling Jiang, Shuang Li, He Huang, Xian Xu
Lycopene plays a crucial role in the biosynthesis pathway of 2-methyl-derythritol-4-phosphate (MEP) and mevalonic acid (MVA). It is a representative product of isoprenoid family, and a typical product of multi-enzyme catalytic reaction in organism. In this paper, we first introduced the general regulation methods in multi-enzyme synthesis reaction, including the construction of multi gene co-expression plasmid, gene order regulation, promoter and ribosome binding site regulation, gene knockout and replacement, aiming at the optimization strategies of multi-enzyme catalytic reaction in lycopene synthesis pathway...
April 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28919886/dna-adenine-methyltransferase-dam-overexpression-impairs-photorhabdus-luminescens-motility-and-virulence
#10
Amaury Payelleville, Anne Lanois, Marie Gislard, Emeric Dubois, David Roche, Stéphane Cruveiller, Alain Givaudan, Julien Brillard
Dam, the most described bacterial DNA-methyltransferase, is widespread in gamma-proteobacteria. Dam DNA methylation can play a role in various genes expression and is involved in pathogenicity of several bacterial species. The purpose of this study was to determine the role played by the dam ortholog identified in the entomopathogenic bacterium Photorhabdus luminescens. Complementation assays of an Escherichia coli dam mutant showed the restoration of the DNA methylation state of the parental strain. Overexpression of dam in P...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28919881/genomic-and-transcriptomic-analysis-of-growth-supporting-dehalogenation-of-chlorinated-methanes-in-methylobacterium
#11
Pauline Chaignaud, Bruno Maucourt, Marion Weiman, Adriana Alberti, Steffen Kolb, Stéphane Cruveiller, Stéphane Vuilleumier, Françoise Bringel
Bacterial adaptation to growth with toxic halogenated chemicals was explored in the context of methylotrophic metabolism of Methylobacterium extorquens, by comparing strains CM4 and DM4, which show robust growth with chloromethane and dichloromethane, respectively. Dehalogenation of chlorinated methanes initiates growth-supporting degradation, with intracellular release of protons and chloride ions in both cases. The core, variable and strain-specific genomes of strains CM4 and DM4 were defined by comparison with genomes of non-dechlorinating strains...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28919459/genome-content-metabolic-pathways-and-biotechnological-potential-of-the-psychrophilic-arctic-bacterium-psychrobacter-sp-dab_al43b-a-source-and-a-host-of-novel-psychrobacter-specific-vectors
#12
Robert Lasek, Lukasz Dziewit, Anna Ciok, Przemyslaw Decewicz, Krzysztof Romaniuk, Zuzanna Jedrys, Daniel Wibberg, Andreas Schlüter, Alfred Pühler, Dariusz Bartosik
Psychrobacter sp. DAB_AL43B, isolated from ornithogenic soil collected on the Arctic island of Spitsbergen, is a newly sequenced psychrophilic strain susceptible to conjugation and electrotransformation. Its genome consists of a circular chromosome (3.3 Mb) and four plasmids (4.4-6.4kb). In silico genome mining and microarray-based phenotypic analysis were performed to describe the metabolic potential of this strain and identify possible biotechnological applications. Metabolic reconstruction indicated that DAB_AL43B prefers low-molecular-weight carboxylates and amino acids as carbon and energy sources...
September 14, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28919348/prevalence-of-colistin-resistance-in-clinical-isolates-of-enterobacteriaceae-a-four-year-cross-sectional-study
#13
Núria Prim, Miquel Turbau, Alba Rivera, Judith Rodríguez-Navarro, Pere Coll, Beatriz Mirelis
Objectives The objectives were to determine the prevalence of colistin resistance in clinical isolates of Enterobacteriaceae, and to gain knowledge on the epidemiological and clinical features of the patients. Methods All colistin-resistant Enterobacteriaceae consecutively isolated from clinical samples in our institution from 2012 to 2015, were included in this cross-sectional study. Intrinsic-resistant species with were excluded. Minimum inhibitory concentration was performed by gradient diffusion. Detection of plasmid-encoded colistin resistance genes mcr-1 and mcr-2 was performed by amplification...
September 14, 2017: Journal of Infection
https://www.readbyqxmd.com/read/28919197/imported-poultry-meat-as-a-source-of-extended-spectrum-cephalosporin-resistant-cmy-2-producing-salmonella-heidelberg-and-s-minnesota-in-european-union-2014-2015
#14
Joana Campos, Joana Mourão, Leonor Silveira, Margarida Saraiva, Cristina Belo Correia, Ana Paula Maçãs, Luísa Peixe, Patrícia Antunes
Extended-spectrum cephalosporin (ESC)-resistant Salmonella has been described at low level in EU, nevertheless the increasing importation of poultry meat could be an important source of epidemic strains carrying ESC-resistant genes. We evaluated ESC resistance and characterized genetic platforms as well as the clonal relatedness of Salmonella isolates from poultry meat products imported into Portugal. All Salmonella isolates recovered from samples of fresh meat destined to be imported into the EU in the scope of Portuguese official border control between 2014 and 2015 were studied...
September 14, 2017: International Journal of Antimicrobial Agents
https://www.readbyqxmd.com/read/28919196/inca-c-plasmids-conferring-high-azithromycin-resistance-in-vibrio-cholera
#15
Ruibai Wang, Haican Liu, Xiuqin Zhao, Jie Li, Kanglin Wan
Azithromycin (AZM) is a clinically important antibiotic to Vibrio cholerae especially for inhibiting colonization of V. cholerae in intestine and the treatment of severe cholera in children and pregnant women. One IncA/C plasmid was isolated from both of the high MIC AZM resistant V cholerae strains of the two mainly pathogenic serogroups, O1 and O139. In the 172 predicted ORFs, 16 genes were related to antibiotic resistance in which 5 were well-defined genes associated with macrolide resistance. The 5 macrolide resistant genes disturbed in two clusters, mphR-mrx-mph(K) and mel-mph2, flanking by insertion sequence (IS) and involving two kinds of mechanisms...
September 14, 2017: International Journal of Antimicrobial Agents
https://www.readbyqxmd.com/read/28919044/susceptibility-of-colistin-resistant-pathogens-to-predatory-bacteria
#16
Sonal Dharani, Dong Hyun Kim, Robert M Q Shanks, Yohei Doi, Daniel E Kadouri
The increase in multidrug-resistant Gram-negative bacterial infections has forced the reintroduction of antibiotics such as colistin. However, the spread of plasmid-borne mcr-1 colistin resistance genes have moved us closer to an era of untreatable Gram-negative infections. To evaluate whether predatory bacteria could be used as a potential therapeutic to treat this upcoming threat, the ability of Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus to prey on several clinically relevant mcr-1-positive, colistin-resistant isolates was evaluated...
September 14, 2017: Research in Microbiology
https://www.readbyqxmd.com/read/28918634/an-engineered-probiotic-for-the-inhibition-of-salmonella-via-tetrathionate-induced-production-of-microcin-h47
#17
Jacob D Palmer, Emma Piattelli, Beth McCormick, Mark Silby, Christopher Brigham, Vanni Bucci
Complications arising from antibiotic-resistant bacteria are becoming one of the key issues in modern medicine. Members of drug-resistant Enterobacteriaceae spp. include opportunistic pathogens (e.g. Salmonella spp.) that are among the leading causes of morbidity and mortality worldwide. Overgrowth of these bacteria is considered a hallmark of intestinal dysbiosis. Microcins (small antimicrobial peptides) produced by some gut commensals can potentially cure these conditions by inhibiting these pathogens, and have been proposed as a viable alternative to antibiotic treatment...
September 17, 2017: ACS Infectious Diseases
https://www.readbyqxmd.com/read/28918285/controlling-cell-volume-for-efficient-phb-production-by-halomonas
#18
Xiao-Ran Jiang, Zhi-Hao Yao, Guo-Qiang Chen
Bacterial morphology is decided by cytoskeleton protein MreB and cell division protein FtsZ encoded by essential genes mreB and ftsZ, respectively. Inactivating mreB and ftsZ lead to increasing cell sizes and cell lengths, respectively, yet seriously reduce cell growth ability. Here we develop a temperature-responsible plasmid expression system for compensated expression of relevant gene(s) in mreB or ftsZ disrupted recombinants H. campaniensis LS21, allowing mreB or ftsZ disrupted recombinants to grow normally at 30°C in a bioreactor for 12h so that a certain cell density can be reached, followed by 36h cell size expansions or cell shape elongations at elevated 37°C at which the mreB and ftsZ encoded plasmid pTKmf failed to replicate in the recombinants and thus lost themselves...
September 13, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28918058/reducible-peg-pod-dna-nanoparticles-for-gene-transfer-in%C3%A2-vitro-and-in%C3%A2-vivo-application-in-a-mouse-model-of-age-related-macular-degeneration
#19
Bhanu Chandar Dasari, Siobhan M Cashman, Rajendra Kumar-Singh
Non-viral gene delivery systems are being developed to address limitations of viral gene delivery. Many of these non-viral systems are modeled on the properties of viruses including cell surface binding, endocytosis, endosomal escape, and nuclear targeting. Most non-viral gene transfer systems exhibit little correlation between in vitro and in vivo efficiency, hampering a systematic approach to their development. Previously, we have described a 3.5 kDa peptide (peptide for ocular delivery [POD]) that targets cell surface sialic acid...
September 15, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28918040/codon-optimized-p1a-encoding-dna-vaccine-toward-a-therapeutic-vaccination-against-p815-mastocytoma
#20
Alessandra Lopes, Kevin Vanvarenberg, Véronique Préat, Gaëlle Vandermeulen
DNA vaccine can be modified to increase protein production and modulate immune response. To enhance the efficiency of a P815 mastocytoma DNA vaccine, the P1A gene sequence was optimized by substituting specific codons with synonymous ones while modulating the number of CpG motifs. The P815A murine antigen production was increased with codon-optimized plasmids. The number of CpG motifs within the P1A gene sequence modulated the immunogenicity by inducing a local increase in the cytokines involved in innate immunity...
September 15, 2017: Molecular Therapy. Nucleic Acids
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