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https://www.readbyqxmd.com/read/29331350/construction-of-pogoduet-an-inducible-bicistronic-vector-for-synthesis-of-recombinant-proteins-in-corynebacterium-glutamicum
#1
Oliver Goldbeck, Gerd M Seibold
The Gram-positive Corynebacterium glutamicum is widely known for its application in the industrial production of amino acids and as a non-pathogenic model organism for cell wall biosynthesis in the group of CMN bacteria. For biotechnological and physiological studies often co-expression of recombinant genes is required, however for C. glutamicum no vector for the independent co-expression of two genes was described. We here created the novel expression vector pOGOduet for C. glutamicum, which carries the ColE1 replicon of E...
January 10, 2018: Plasmid
https://www.readbyqxmd.com/read/29327233/dcaf1-is-involved-in-hcv-replication-through-regulation-of-mir-122
#2
Yanling Yan, Conghui Li, Binlian Sun, Rongge Yang
Hepatitis C virus (HCV) is a worldwide threaten to human health with a high ratio of chronic infections. Recently, we found that Vpr-mediated regulation of HCV replication depends on the host protein DDB1-Cul4 associate factor 1 (DCAF1), implying that DCAF1 might be involved in the replication of HCV. In this study, we demonstrated that DCAF1 knockdown reduced HCV replication both in the infectious (JFH1) and replicon (Con1) systems. Further investigation showed a negative regulation of HCV internal ribosome entry site (IRES)-mediated translation by DCAF1...
January 11, 2018: Archives of Virology
https://www.readbyqxmd.com/read/29322851/clinical-and-virological-implications-of-a1846t-and-c1913a-g-mutations-of-hepatitis-b-virus-genome-in-severe-liver-diseases
#3
Hong Zang, Zhihui Xu, Yan Liu, Xiaodong Li, Yihui Rong, Ling Jiang, Shaoli You, Jinhua Hu, Jun Zhao, Dongping Xu, Shaojie Xin
OBJECTIVE: Mutations occurring within different genes of hepatitis B virus (HBV) genome may have different clinical implications. This study aimed to observe the clinical and virological implications of the A1846T and C1913A/G mutations of HBV genome in the development and treatment outcome of severe liver diseases, which has not been previously determined. MATERIALS AND METHODS: A total of 438 cases of patients with liver diseases were retrospectively reviewed, including 146 with mild chronic hepatitis B infection (CHB-M), 146 with severe chronic hepatitis B infection (CHB-S), and 146 with acute-on-chronic liver failure (ACLF)...
January 11, 2018: Scandinavian Journal of Gastroenterology
https://www.readbyqxmd.com/read/29321328/activities-of-thrombin-and-factor-xa-are-essential-for-replication-of-hepatitis-e-virus-and-are-possibly-implicated-in-the-orf1-polyprotein-processing
#4
Gayatri D Kanade, Kunal D Pingale, Yogesh A Karpe
Hepatitis E virus (HEV) is a clinically important positive-sense RNA virus. The ORF1 of HEV encodes a non-structural polyprotein of 1693 amino acids. It is not clear whether the ORF1 polyprotein is processed into distinct enzymatic domains. Many researchers have attempted to understand the mechanisms of pORF1 processing. However, these studies gave varied results and could never convincingly establish the mechanism of pORF1 processing. In this study, we demonstrated the possible role of thrombin and factor Xa in pORF1 processing...
January 10, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29305512/molecular-epidemiology-of-dairy-cattle-associated-escherichia-coli-carrying-blactx-m-genes-in-washington-state
#5
Josephine A Afema, Sara Ahmed, Thomas E Besser, Lisa P Jones, William M Sischo, Margaret A Davis
An increase in prevalence of commensal Escherichia coli carrying blaCTX-M genes among dairy cattle was observed between 2008 and 2012 in Washington State. To study the molecular epidemiology of this change, we selected 126 blaCTX-M-positive and 126 blaCTX-M-negative isolates for multi-locus sequence typing (MLST) and antibiotic resistance phenotyping from E. coli obtained during a previous study. For 99 isolates we also determined the blaCTX-M alleles using PCR and sequencing and identified the replicon types of blaCTX-M carrying plasmids...
January 5, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29303020/risk-factors-outcomes-and-genotypes-of-carbapenem-nonsusceptible-klebsiella-pneumoniae-bloodstream-infection-a-three-year-retrospective-study-in-a-large-tertiary-hospital-in-northern-china
#6
Si-Han Zheng, Sheng-Ji Cao, Hui Xu, Dan Feng, Li-Ping Wan, Guang-Jie Wang, Xiao-Guang Xiao
OBJECTIVE: To investigate the independent risk factors, outcomes and genotypes associated with carbapenem-non-susceptible K. pneumoniae bloodstream infections (BSIs) in northern China from 2014 to 2016. METHODS: Over a three-year period, a total of 289 K. pneumoniae BSI patients were identified. Medical records were extracted to obtain the clinical information. Polymerase chain reactions (PCRs) were performed to analyse the multilocus sequence typing (MLST) types, Klebsiella pneumoniae carbapenemase (KPC) and metallo-β-lactamases (MBL) genes, for replicon typing of the 10 randomly selected carbapenem-non-susceptible K...
January 5, 2018: Infectious Diseases
https://www.readbyqxmd.com/read/29288699/development-of-a-replicon-cell-line-based-high-throughput-antiviral-assay-for-screening-inhibitors-of-zika-virus
#7
Jia-Qi Li, Cheng-Lin Deng, Dayong Gu, Xiao Li, Lei Shi, Jian'an He, Qiu-Yan Zhang, Bo Zhang, Han-Qing Ye
Zika virus (ZIKV) is an important emerging human pathogen associated with microcephaly, Guillain-Barré syndrome and meningoencephalitis. Developing rapid and reliable HTS assay is important for ZIKV drug discovery. Here, we constructed a dicistronic ZIKV replicon (ZIKV-Pac-Rluc-Rep) that contained the Renilla luciferase (Rluc) reporter gene separated from the puromycin N-acetyl-transferase (Pac) selectable marker by a short peptide cleavage site. A clonal replicon cell line stably expressing high level of ZIKV replicon was established by selection with puromycin...
December 27, 2017: Antiviral Research
https://www.readbyqxmd.com/read/29287681/alphavirus-vector-based-replicon-particles-expressing-multivalent-cross-protective-lassa-virus-glycoproteins
#8
Min Wang, Jenny Jokinen, Irina Tretyakova, Peter Pushko, Igor S Lukashevich
Lassa virus (LASV) is the most prevalent rodent-borne arenavirus circulated in West Africa. With population at risk from Senegal to Nigeria, LASV causes Lassa fever and is responsible for thousands of deaths annually. High genetic diversity of LASV is one of the challenges for vaccine R&D. We developed multivalent virus-like particle vectors (VLPVs) derived from the human Venezuelan equine encephalitis TC-83 IND vaccine (VEEV) as the next generation of alphavirus-based bicistronic RNA replicon particles. The genes encoding VEEV structural proteins were replaced with LASV glycoproteins (GPC) from distantly related clades I and IV with individual 26S promoters...
December 26, 2017: Vaccine
https://www.readbyqxmd.com/read/29282569/emergence-and-establishment-of-kpc-2-producing-st11-klebsiella-pneumoniae-in-a-general-hospital-in-shanghai-china
#9
Jingxian Liu, Jing Yu, Feng Chen, Jiajia Yu, Patricia Simner, Pranita Tamma, Ying Liu, Lisong Shen
The aim of this study was to investigate the characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP) collected during an outbreak in a Chinese teaching hospital and to provide insights into the prevention and control of nosocomial infection. We collected unique CRKP clinical isolates from 2009 to 2013. Antibiotic-resistant genes were identified by polymerase chain reaction (PCR) and sequencing. The isolates were typed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)...
December 27, 2017: European Journal of Clinical Microbiology & Infectious Diseases
https://www.readbyqxmd.com/read/29279444/detection-of-escherichia-coli-producing-ctx-m-1-group-extended-spectrum-%C3%AE-lactamases-from-pigs-in-aichi-prefecture-japan-between-2015-and-2016
#10
Chihiro Norizuki, Kumiko Kawamura, Jun-Ichi Wachino, Masahiro Suzuki, Noriyuki Nagano, Takaaki Kondo, Yoshichika Arakawa
We investigated the prevalence and characteristics of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolates from Japanese pigs. A total of 345 pig fecal specimens were collected from 30 farms in Aichi prefecture, Japan, between June 2015 and April 2016, and 22 unique ESBL-producing E. coli were isolated from 16 samples of eight farms. The ESBL types included CTX-M-15 (54.5%), CTX-M-55 (27.2%), CTX-M-3 (0.9%), and CTX-M-14 (0.9%). The predominant plasmid replicon type was IncN, and they carried blaCTX-M-55...
December 26, 2017: Japanese Journal of Infectious Diseases
https://www.readbyqxmd.com/read/29250051/successful-establishment-of-plasmids-r1-and-pmv158-in-a-new-host-requires-the-relief-of-the-transcriptional-repression-of-their-essential-rep-genes
#11
José Á Ruiz-Masó, Luis M Luengo, Inmaculada Moreno-Córdoba, Ramón Díaz-Orejas, Gloria Del Solar
Although differing in size, encoded traits, host range, and replication mechanism, both narrow-host-range theta-type conjugative enterobacterial plasmid R1 and promiscuous rolling-circle-type mobilizable streptococcal plasmid pMV158 encode a transcriptional repressor protein, namely CopB in R1 and CopG in pMV158, involved in replication control. The gene encoding CopB or CopG is cotranscribed with a downstream gene that encodes the replication initiator Rep protein of the corresponding plasmid. However, whereas CopG is an auto-repressor that inhibits transcription of the entire copG-repB operon, CopB is expressed constitutively and represses a second, downstream promoter that directs transcription of repA...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29246842/a-helicase-containing-module-defines-a-family-of-pcd630-like-plasmids-in-clostridium-difficile
#12
Wiep Klaas Smits, J Scott Weese, Adam P Roberts, Céline Harmanus, Bastian Hornung
Clostridium difficile is a Gram-positive and sporulating enteropathogen that is a major cause of healthcare-associated infections. Even though a large number of genomes of this species have been sequenced, only a few plasmids have been described in the literature. Here, we use a combination of in silico analyses and laboratory experiments to show that plasmids are common in C. difficile. We focus on a group of plasmids that share similarity with the plasmid pCD630, from the reference strain 630. The family of pCD630-like plasmids is defined by the presence of a conserved putative helicase that is likely part of the plasmid replicon...
December 12, 2017: Anaerobe
https://www.readbyqxmd.com/read/29246301/profiling-the-mismatch-tolerance-of-argonaute-2-through-deep-sequencing-of-sliced-polymorphic-viral-rnas
#13
Pantazis I Theotokis, Louise Usher, Christopher K Kortschak, Ed Schwalbe, Sterghios A Moschos
Low allelic and clonal variability among endogenous RNAi targets has focused mismatch tolerance studies to RNAi-active guide strands. However, the inherent genomic instability of RNA viruses such as hepatitis C virus (HCV) gives rise to quasi-species mutants within discrete clones: this facilitates mismatch tolerance studies from a target perspective. We recently quantified the slicing imprecision of Argonaute 2 using small interfering RNA (siRNA) analogs of the DNA-directed RNAi drug TT-034 and next-generation sequencing of 5' RNA ligase-mediated rapid amplification of cDNA ends (RACE-SEQ)...
December 15, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/29243837/a-minireplicon-of-plasmid-pbmb26-represents-a-new-typical-replicon-in-the-megaplasmids-of-bacillus-cereus-group
#14
Pengxia Wang, Yiguang Zhu, Hui Shang, Yun Deng, Ming Sun
A new minireplicon (rep26 minireplicon) from pBMB26, the 188 kb indigenous plasmid related to spore-crystal association (SCA) phenotype in Bacillus thuringiensis strain YBT-020, was characterized. A 12 kb EcoRI fragment, which encoded 10 putative open reading frames (ORFs), was capable of supporting replication when cloned in a replication probe vector. Deletion and frame shift mutation analysis showed that a 4.1 kb region encompassing two putative ORFs (orf21 and orf22) was essential for the plasmid replication in B...
December 15, 2017: Journal of Basic Microbiology
https://www.readbyqxmd.com/read/29242561/hepatitis-b-virus-persistence-in-mice-reveals-il-21-and-il-33-as-regulators-of-viral-clearance
#15
Zhongliang Shen, Huijuan Yang, Sisi Yang, Wei Wang, Xiaoxian Cui, Xian Zhou, Wei Liu, Shaokun Pan, Yanfeng Liu, Junqi Zhang, Jiming Zhang, Youhua Xie, Jing Liu
Hepatitis B virus (HBV) generally causes self-limiting infection in immunocompetent adults, but establishes chronic infection in some adults and in most maternally infected infants. Factors determining clearance versus persistence are not fully understood. Hydrodynamic injection (HDI) of HBV replicon plasmid via tail vein generally results in quick clearance in immunocompetent adult mice. Here, we report the identification of strain-specific persistence of HBV in mice: one genotype B strain, designated BPS, persisted up to 33 weeks in ~50% of HDI mice...
December 14, 2017: Nature Communications
https://www.readbyqxmd.com/read/29237842/human-norovirus-ns3-has-rna-helicase-and-chaperoning-activities
#16
Teng-Feng Li, Myra Hosmillo, Hella Schwanke, Ting Shu, Zhaowei Wang, Lei Yin, Stephen Curry, Ian G Goodfellow, Xi Zhou
RNA remodeling proteins, including RNA helicases and chaperones, act to remodel RNA structures and/or protein-RNA interactions, and are required for all processes involving RNAs. Although many viruses encode RNA helicases and chaperones, their in vitro activities and their roles in infected cells largely remain elusive. Noroviruses are a diverse group of positive-stranded RNA viruses in the family Caliciviridae, and constitute a significant and potentially fatal threat to human health. Here we report that protein NS3 encoded by human norovirus has both ATP-dependent RNA helicase activity that unwinds RNA helices and ATP-independent RNA chaperoning activity that can remodel structured RNAs and facilitate strand-annealing...
December 13, 2017: Journal of Virology
https://www.readbyqxmd.com/read/29237086/enterobacteriaceae-producing-oxa-48-like-carbapenemases-in-poland-2013-january-2017
#17
R Izdebski, A Baraniak, D Zabicka, M Machulska, P Urbanowicz, J Fiett, E Literacka, K Bojarska, A Kozinska, B Zieniuk, W Hryniewicz, M Gniadkowski
Objectives: To analyse OXA-48 (OXA-48/181)-type carbapenemase-producing Enterobacteriaceae reported in Poland from 2013 until January 2017. Methods: Bacterial isolates were typed by PFGE and MLST. Genes coding for OXA-48/181 types and other β-lactamases were amplified and sequenced. Mobile elements with blaOXA-48/181-like genes were PCR mapped. blaOXA-48/181-carrying plasmids were characterized by nuclease S1-hybridization profiling, transfer assays and PCR-based replicon typing, while the chromosomal location of the genes was confirmed by the I-CeuI analysis...
December 9, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29237003/illumina-short-read-and-minion-long-read-wgs-to-characterize-the-molecular-epidemiology-of-an-ndm-1-serratia-marcescens-outbreak-in-romania
#18
H T T Phan, N Stoesser, I E Maciuca, F Toma, E Szekely, M Flonta, A T M Hubbard, L Pankhurst, T Do, T E A Peto, A S Walker, D W Crook, D Timofte
Background and Objectives: Serratia marcescens is an emerging nosocomial pathogen, and the carbapenemase blaNDM has been reported in several surveys in Romania. We aimed to investigate the molecular epidemiology of S. marcescens in two Romanian hospitals over 2010-15, including a neonatal NDM-1 S. marcescens outbreak. Methods: Isolates were sequenced using Illumina technology together with carbapenem-non-susceptible NDM-1-positive and NDM-1-negative Klebsiella pneumoniae and Enterobacter cloacae to provide genomic context...
December 8, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29235977/proprotein-convertase-subtilisin-kexin-type-9-inhibits-hepatitis-c-virus-replication-through-interacting-with-ns5a
#19
Zhubing Li, Qiang Liu
Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a serine protease actively involved in regulating lipid homeostasis. Although PCSK9 has been shown to inhibit hepatitis C virus (HCV) entry and replication, the underlying mechanisms have not been thoroughly characterized. Moreover, whether PCSK9 regulates HCV translation and assembly/secretion has not been determined. We therefore further studied the effects of PCSK9 on the HCV life cycle. We showed that PCSK9 did not affect HCV translation or assembly/secretion...
December 13, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/29229976/a-novel-tn1696-like-composite-transposon-tn6404-harboring-bla-imp-4-in-a-klebsiella-pneumoniae-isolate-carrying-a-rare-esbl-gene-bla-sfo-1
#20
Kai Zhou, Wei Yu, Ping Shen, Haifeng Lu, Baohong Wang, John W A Rossen, Yonghong Xiao
Genetic determinants of a clinical Klebsiella pneumoniae isolate (KP1814) coproducing IMP-4 and a rare ESBL gene SFO-1 was investigated. KP1814 belongs to a novel sequence type (ST) assigned to ST2270. WGS identified four circular DNA sequences in KP1814, including two multidrug-resistance (MDR) plasmids, one virulence plasmid, and one circular form. The MDR plasmid pKP1814-1 (299.9 Kb) is untypeable, and carries two large mosaic multiresistance regions (MRRs). bla SFO-1 and bla IMP-4 co-exists on MRR1, and bla SFO-1 is associated with an IS/Tn-independent genetic context...
December 11, 2017: Scientific Reports
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