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https://www.readbyqxmd.com/read/28746401/identification-of-one-b-cell-epitope-from-ns1-protein-of-duck-tembusu-virus-with-monoclonal-antibodies
#1
Jinfeng Ti, Zhijie Li, Xiuli Li, Yunjian Lu, Youxiang Diao, Fang Li
This study describes the identification of one linear B-cell epitope on TMUV NS1 protein with monoclonal antibody (mAb) 3G2 by indirect enzyme-linked immunosorbent assay (ELISA). In this study, NS1 protein was expressed in prokaryotic expression system and purified. One mAb against NS1 protein was generated from Balb/c mice immunized with recombinant protein NS1. A set of 35 partially-overlapping polypeptides covering the entire NS1 protein was expressed with PGEX-6P-1 vector and screened with mAb 3G2. One polypeptide against the mAb was acquired and identified by indirect ELISA and western-blot...
2017: PloS One
https://www.readbyqxmd.com/read/28744694/structural-and-functional-diversity-of-caspase-homologues-in-non-metazoan-organisms
#2
REVIEW
Marina Klemenčič, Christiane Funk
Caspases, the proteases involved in initiation and execution of metazoan programmed cell death, are only present in animals, while their structural homologues can be found in all domains of life, spanning from simple prokaryotes (orthocaspases) to yeast and plants (metacaspases). All members of this wide protease family contain the p20 domain, which harbours the catalytic dyad formed by the two amino acid residues, histidine and cysteine. Despite the high structural similarity of the p20 domain, metacaspases and orthocaspases were found to exhibit different substrate specificities than caspases...
July 25, 2017: Protoplasma
https://www.readbyqxmd.com/read/28743817/bacterial-signaling-nucleotides-inhibit-yeast-cell-growth-by-impacting-mitochondrial-and-other-specifically-eukaryotic-functions
#3
Andy Hesketh, Marta Vergnano, Chris Wan, Stephen G Oliver
We have engineered Saccharomyces cerevisiae to inducibly synthesize the prokaryotic signaling nucleotides cyclic di-GMP (cdiGMP), cdiAMP, and ppGpp in order to characterize the range of effects these nucleotides exert on eukaryotic cell function during bacterial pathogenesis. Synthetic genetic array (SGA) and transcriptome analyses indicated that, while these compounds elicit some common reactions in yeast, there are also complex and distinctive responses to each of the three nucleotides. All three are capable of inhibiting eukaryotic cell growth, with the guanine nucleotides exhibiting stronger effects than cdiAMP...
July 25, 2017: MBio
https://www.readbyqxmd.com/read/28736192/the-presence-of-phb-granules-in-cytoplasm-protects-non-halophilic-bacterial-cells-against-the-harmful-impact-of-hypertonic-environments
#4
Stanislav Obruca, Petr Sedlacek, Filip Mravec, Vladislav Krzyzanek, Jana Nebesarova, Ota Samek, Dan Kucera, Pavla Benesova, Kamila Hrubanova, Miluse Milerova, Ivana Marova
Numerous prokaryotes accumulate polyhydroxybutyrate (PHB) intracellularly as a storage material. It has also been proposed that PHB accumulation improves bacterial stress resistance. Cupriavidus necator and its PHB non-accumulating mutant were employed to investigate the protective role of PHB under hypertonic conditions. The presence of PHB granules enhanced survival of the bacteria after exposure to hypertonic conditions. Surprisingly, when coping with such conditions, the bacteria did not utilize PHB to harvest carbon or energy, suggesting that, in the osmotic upshock of C...
July 20, 2017: New Biotechnology
https://www.readbyqxmd.com/read/28735080/recombinant-truncated-e-protein-as-a-new-vaccine-candidate-against-nontypeable-h-influenzae-its-expression-and-immunogenic-evaluation
#5
Ava Behrouzi, Saeid Bouzari, Farzam Vaziri, Abolfazl Fateh, Parviz Afrough, Atefeh Davoudi Vijeh Motlagh, Seyed Davar Siadat
Protein E (PE) is a conserved entity observed in both nontypeable Haemophilus influenzae (NTHi) and encapsulated H. influenzae. This is a small surface lipoprotein, consisting of only 160 amino acids, involved in the adhesion of H. influenzae to various types of epithelial cells. A 384-bp-long fragment from NTHi PE was cloned into the prokaryotic expression vector pBAD-gIIIA. The recombinant protein was expressed with arabinose and then purified by affinity purification on an Ni-NTA agarose matrix. BALB/c mice were immunized by subcutaneous injection with purified recombinant truncated PE mixed with an alum adjuvant...
July 19, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28734049/development-of-aptamers-against-unpurified-proteins
#6
Shinichi Goto, Kaori Tsukakoshi, Kazunori Ikebukuro
SELEX (Systematic Evolution of Ligands by EXponential enrichment) has been widely used for the generation of aptamers against target proteins. However, its requirement for pure target proteins remains a major problem in aptamer selection, as procedures for protein purification from crude bio-samples are not only complicated but also time and labour consuming. This is because native proteins can be found in a large number of diverse forms because of posttranslational modifications and their complicated molecular conformations...
July 22, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28730763/dna-mismatch-repair-deficiency-in-lung-and-oral-cavity-carcinomas-the-role-of-histogenetic-origin
#7
Konstantina N Spetsotaki, Evangelos Tsiambas, Athanasios Stamatelopoulos, Panagiotis P Fotiades, Ioannis Kastanioudakis, Pericles Tomos, Vasileios Ragos
DNA mismatch repair system (DNA MMR) is a crucial genetic mechanism for DNA homeostasis in prokaryotic and eukaryotic cells. During DNA replication and also recombination, point intra-nucleotide errors including base deletion, insertion, and mis-incorporation happen. These raised abnormalities in the newly synthesized DNA strand could affect negatively the stability of the molecule and the function of the corresponding genes. DNA MMR proteins prevent these errors by recognizing and repairing them, securing directly the normal anatomy of the DNA double strand and indirectly the expression of the genes...
May 2017: Journal of B.U.ON.: Official Journal of the Balkan Union of Oncology
https://www.readbyqxmd.com/read/28730576/effects-of-boron-supplementation-on-expression-of-hsp70-in-the-spleen-of-african-ostrich
#8
Khaliq Haseeb, Jing Wang, Ke Xiao, Ke-Li Yang, Peng-Peng Sun, Xing-Tong Wu, You Luo, Hui Song, Hua-Zhen Liu, Ju-Ming Zhong, Ke-Mei Peng
Increased synthesis of heat shock protein 70 (Hsp70) occurs in prokaryotes and eukaryotes in response to physiological, environmental, and chemical exposures, thus allowing the cell survival from fatal conditions. Hsp70 cytoprotective properties may be clarified by its anti-apoptotic function. Boron has been reported to play an essential role in various organ developments and metabolisms. However, it is not known if boron is also able to modulate the Hsp70. In the present study, the actions of boron on ostrich spleen and expression level of Hsp70 were investigated...
July 20, 2017: Biological Trace Element Research
https://www.readbyqxmd.com/read/28730447/construction-and-in-vivo-testing-of-prokaryotic-riboregulators
#9
Alexander A Green
RNAs that are transcribed and self-assemble within living cells are valuable tools for regulating and organizing cellular activities. Riboregulators, in particular, have been widely used for modulating translation and transcription in response to cognate transactivating or trigger RNAs, enabling cells to evaluate logic operations and to respond to environmental cues. Herein we detail a set of methods for the rapid construction and testing of prokaryotic riboregulators in Escherichia coli. These methods enable construction of dozens of riboregulator plasmids at the same time without the use of restriction enzymes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28730145/prokaryotic-rna-associated-to-bacterial-viability-induces-polymorphonuclear-neutrophil-activation
#10
Nahuel Rodriguez-Rodrigues, Luis A Castillo, Verónica I Landoni, Daiana Martire-Greco, M Ayelén Milillo, Paula Barrionuevo, Gabriela C Fernández
Polymorphonuclear neutrophils (PMN) are the first cellular line of antibacterial host defense. They sense pathogens through recognition of pathogen-associated molecular patterns (PAMPs) by innate pattern recognition receptors, such as Toll-like receptors (TLR). The aim of this study was to investigate whether PMN sense bacterial viability and explore which viability factor could be involved in this phenomenon. For this purpose, different functions were evaluated in isolated human PMN using live Escherichia coli (Ec) and heat-killed Ec (HK-Ec)...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28729070/abc-transporters-in-the-liver-fluke-opisthorchis-felineus
#11
Viatcheslav A Mordvinov, Nikita I Ershov, Darya S Pirozhkova, Yuri V Pakharukov, Maria Y Pakharukova
ATP-binding cassette transporters (ABC transporters) are essential components of normal cellular physiological machinery in all eukaryotic and prokaryotic species, including parasites. Some ABC transporters, e.g., P-glycoproteins, are involved in the efflux of toxins and xenobiotics from the cell. At present, nothing is known about ABC transporter genes in epidemiologically important liver flukes from the Opisthorchiidae family, including European liver fluke Opisthorchis felineus. Opisthorchiasis caused by O...
July 17, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28723971/termination-factor-rho-from-the-control-of-pervasive-transcription-to-cell-fate-determination-in-bacillus-subtilis
#12
Vladimir Bidnenko, Pierre Nicolas, Aleksandra Grylak-Mielnicka, Olivier Delumeau, Sandrine Auger, Anne Aucouturier, Cyprien Guerin, Repoila Francis, Jacek Bardowski, Stéphane Aymerich, Elena Bidnenko
In eukaryotes, RNA species originating from pervasive transcription are regulators of various cellular processes, from the expression of individual genes to the control of cellular development and oncogenesis. In prokaryotes, the function of pervasive transcription and its output on cell physiology is still unknown. Most bacteria possess termination factor Rho, which represses pervasive, mostly antisense, transcription. Here, we investigate the biological significance of Rho-controlled transcription in the Gram-positive model bacterium Bacillus subtilis...
July 19, 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28717130/polymicrobial-infections-in-brain-tissue-from-alzheimer-s-disease-patients
#13
Diana Pisa, Ruth Alonso, Ana M Fernández-Fernández, Alberto Rábano, Luis Carrasco
Several studies have advanced the idea that the etiology of Alzheimer's disease (AD) could be microbial in origin. In the present study, we tested the possibility that polymicrobial infections exist in tissue from the entorhinal cortex/hippocampus region of patients with AD using immunohistochemistry (confocal laser scanning microscopy) and highly sensitive (nested) PCR. We found no evidence for expression of early (ICP0) or late (ICP5) proteins of herpes simplex virus type 1 (HSV-1) in brain sections. A polyclonal antibody against Borrelia detected structures that appeared not related to spirochetes, but rather to fungi...
July 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28708533/expression-and-molecular-cloning-of-interferon-stimulated-genes-in-buffalo-bubalus-bubalis
#14
Nipuna Thakur, Girjesh Singh, A Paul, J Bharati, G Rajesh, Vidyalakshmi Gm, V S Chouhan, S K Bhure, V P Maurya, G Singh, M Sarkar
Buffalo, the most important livestock species in tropical India, remains to be a poor breeder mainly due to embryonic mortality (65%) occurring mostly between 16 and 18 days of pregnancy. Early and accurate diagnosis of pregnancy can thus become a boon for successful herd management in buffalo. However, most of the currently available methods allow diagnosis only after 30 days post AI. Interferon tau (IFNT), the first pregnancy recognition signal in ruminants is one such molecule, which stimulates expression of various Interferon stimulated genes (ISGs) in the peripheral blood mononuclear cells (PBMC's) concomitant with IFNT signaling which occurs around maternal recognition of pregnancy (MRP)...
June 1, 2017: Theriogenology
https://www.readbyqxmd.com/read/28707664/-a-study-of-recombinant-human-sestrin-1-and-sestrin-2-proteins-produced-in-a-prokaryotic-system
#15
N Rai, R Kumar, Md A Haque, Md I Hassan, S Dey
Sestrins are highly conserved stress-inducible proteins capable of suppressing the production of ROS and signalling through mTORC1. Here we report a study of human sestrin1 (sesn1) and sestrin2 (sesn2) proteins produced in a pET28^(+) vector based prokaryotic system. Mass spectrometry analysis, western blot and surface plasmon resonance (SPR) of affinity purified sesn1 and sesn2 proteins confirmed their identity; biophysical characteristics were observed using circular dichroism (CD) showing that sesn1 and sesn2 have a predominant α-helical structure...
May 2017: Molekuliarnaia Biologiia
https://www.readbyqxmd.com/read/28707598/protein-tagging-destruction-and-infection
#16
Chetana Bhaskarla, Manoj Bhosale, Pip Banerjee, Nagasuma Chandra, Dipankar Nandi
Cells possess protein quality control mechanisms to maintain proper cellular homeostasis. In eukaryotes, the roles of the ubiquitination and proteasome-mediated degradation of cellular proteins is well established. Recent studies have elucidated protein tagging mechanisms in prokaryotes, involving transfer messenger RNA (tmRNA) and pupylation. In this review, newer insights and bioinformatics analysis of two distinct bacterial protein tagging machineries are discussed. The machinery for tmRNA-mediated tagging is present in several eubacterial representatives, e...
July 13, 2017: Current Protein & Peptide Science
https://www.readbyqxmd.com/read/28704355/a-new-view-into-prokaryotic-cell-biology-from-electron-cryotomography
#17
Catherine M Oikonomou, Grant J Jensen
No abstract text is available yet for this article.
February 2017: Nature Reviews. Microbiology
https://www.readbyqxmd.com/read/28701712/use-of-single-frequency-impedance-spectroscopy-to-characterize-the-growth-dynamics-of-biofilm-formation-in-pseudomonas-aeruginosa
#18
Jozef B J H van Duuren, Mathias Müsken, Bianka Karge, Jürgen Tomasch, Christoph Wittmann, Susanne Häussler, Mark Brönstrup
Impedance spectroscopy has been applied in prokaryotic and eukaryotic cytometry as a label-free method for the investigation of adherent cells. In this paper, its use for characterizing the growth dynamics of P. aeruginosa biofilms is described and compared to crystal violet staining and confocal microscopy. The method allows monitoring the growth of biofilm-forming P. aeruginosa in a continuous and label-free manner over a period of 72 h in a 96 well plate format. Impedance curves obtained for P. aeruginosa PA14 wild type and mutant strains with a transposon insertion in pqsA and pelA genes exhibited distinct phases...
July 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28696276/progress-in-our-understanding-of-wzx-flippase-for-translocation-of-bacterial-membrane-lipid-linked-oligosaccharide
#19
Yaoqin Hong, Michael A Liu, Peter R Reeves
Translocation of lipid-linked oligosaccharides is a common theme across prokaryotes and eukaryotes. For bacteria, such activity is used in cell wall construction, polysaccharide synthesis and the relatively recently discovered protein glycosylation. To the best of our knowledge, the Gram-negative inner-membrane flippase Wzx was the first protein identified as being involved in oligosaccharide translocation, and yet we still have only a limited understanding of this protein after three decades of research. At present, Wzx is known to be a multi-transmembrane protein with enormous sequence diversity that flips oligosaccharide substrates with varying degrees of preference...
July 10, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28689775/gene-characteristics-immune-and-stress-responses-of-pmprx1-in-black-tiger-shrimp-penaeus-monodon-insights-from-exposure-to-pathogenic-bacteria-and-toxic-environmental-stressors
#20
Ruiqian Bu, Pengfei Wang, Chao Zhao, Weiyang Bao, Lihua Qiu
Peroxiredoxins (Prxs) are ubiquitous, multifunctional and evolutionarily conserved enzymes that can protect cells from oxidative damage caused by ROS and play a vital role in immune responses. Here, a full-length Prx1 cDNA sequence (PmPrx1) was isolated from Penaeus monodon. The PmPrx1 cDNA was 951 base pairs (bp), encoding 198 amino acid polypeptides. The results of qRT-PCR showed that the PmPrx1 mRNA was ubiquitously expressed in all tissues tested and had a comparatively high expression level in immune-associated tissues (gill, hepatopancreas)...
July 6, 2017: Developmental and Comparative Immunology
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