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Heterologous expression lab

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https://www.readbyqxmd.com/read/29621349/signature-pathway-expression-of-xylose-utilization-in-the-genetically-engineered-industrial-yeast-saccharomyces-cerevisiae
#1
Quanzhou Feng, Z Lewis Liu, Scott A Weber, Shizhong Li
Haploid laboratory strains of Saccharomyces cerevisiae are commonly used for genetic engineering to enable their xylose utilization but little is known about the industrial yeast which is often recognized as diploid and as well as haploid and tetraploid. Here we report three unique signature pathway expression patterns and gene interactions in the centre metabolic pathways that signify xylose utilization of genetically engineered industrial yeast S. cerevisiae NRRL Y-50463, a diploid yeast. Quantitative expression analysis revealed outstanding high levels of constitutive expression of YXI, a synthesized yeast codon-optimized xylose isomerase gene integrated into chromosome XV of strain Y-50463...
2018: PloS One
https://www.readbyqxmd.com/read/29545204/baculovirus-driven-protein-expression-in-insect-cells-a-benchmarking-study
#2
Peggy Stolt-Bergner, Christian Benda, Tim Bergbrede, Hüseyin Besir, Patrick H N Celie, Cindy Chang, David Drechsel, Ariane Fischer, Arie Geerlof, Barbara Giabbai, Joop van den Heuvel, Georg Huber, Wolfgang Knecht, Anita Lehner, Regis Lemaitre, Kristina Nordén, Gwynn Pardee, Ines Racke, Kim Remans, Astrid Sander, Judith Scholz, Magda Stadnik, Paola Storici, Daniel Weinbruch, Isabel Zaror, Linda H L Lua, Sabine Suppmann
Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. The availability of robust insect cell lines, serum-free media, a range of vectors and commercially-packaged kits have supported the demand for maximizing the exploitation of the baculovirus-insect cell expression system. Naturally, this resulted in varied strategies adopted by different laboratories to optimize protein production. Most laboratories have preference in using either the E...
March 12, 2018: Journal of Structural Biology
https://www.readbyqxmd.com/read/29540599/development-of-a-broadly-accessible-vee-replicon-particle-vaccine-platform
#3
Sudhakar Agnihothram, Vineet D Menachery, Boyd L Yount, Lisa C Lindesmith, Trevor Scobey, Alan Whitmore, Alexandra Schäfer, Mark T Heise, Ralph S Baric
Zoonotic viruses circulate as swarms in animal reservoirs and can emerge into human populations causing epidemics that adversely affect public health. Portable, safe, and effective vaccine platforms are needed in the context of these outbreak and emergence situations. In this manuscript, we report the generation and characterization of an alphavirus replicon vaccine platform based on a non-select agent, attenuated Venezuelan equine encephalitis (VEE) virus vaccine strain 3526 (VRP 3526). Using both noroviruses and coronaviruses as model systems, we demonstrate utility of the VRP 3526 platform in generation of recombinant proteins, production of virus like particles, and in vivo efficacy as a vaccine against emergent viruses...
March 14, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29535210/rapid-production-of-pure-recombinant-actin-isoforms-in-pichia-pastoris
#4
Tomoyuki Hatano, Salvatore Alioto, Emanuele Roscioli, Saravanan Palani, Scott T Clarke, Anton Kamnev, Juan Ramon Hernandez-Fernaud, Lavanya Sivashanmugam, Bernardo Chapa-Y-Lazo, Alexandra M E Jones, Robert C Robinson, Karuna Sampath, Masanori Mishima, Andrew D McAinsh, Bruce L Goode, Mohan K Balasubramanian
Actins are major eukaryotic cytoskeletal proteins, which perform many important cell functions, including cell division, cell polarity, wound healing, and muscle contraction. Despite obvious drawbacks, muscle actin, which is easily purified, is used extensively presently for biochemical studies of actin cytoskeleton from other organisms / cell types. Here we report a rapid and cost-effective method to purify heterologous actins expressed in the yeast Pichia pastoris Actin is expressed as a fusion with the actin-binding protein thymosin β4 and purified using an affinity tag introduced in the fusion...
March 13, 2018: Journal of Cell Science
https://www.readbyqxmd.com/read/29455388/heterologous-expression-of-rtshyal-1-the-first-recombinant-hyaluronidase-of-scorpion-venom-produced-in-pichia-pastoris-system
#5
Fernanda Gobbi Amorim, Johara Boldrini-França, Karla de Castro Figueiredo Bordon, Iara Aimê Cardoso, Edwin De Pauw, Loïc Quinton, Simone Kashima, Eliane Candiani Arantes
In general, hyaluronidases have a broad potential application on medicine and esthetics fields. Hyaluronidases from animal venoms cleave hyaluronan present in the extracellular matrix, acting as spreading factors of toxins into the tissues of the victim. However, the in-depth characterization of hyaluronidase from animal venoms has been neglected due to its instability and low concentration in the venom, which hamper its isolation. Thus, heterologous expression of hyaluronidase acts as a biotechnological tool in the obtainment of enough amounts of the enzyme for structural and functional studies...
February 17, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29438403/release-of-transcriptional-repression-through-the-hcr-promoter-region-confers-uniform-expression-of-hwp1-on-surfaces-of-candida-albicans-germ-tubes
#6
Samin Kim, Quoc Bao Nguyen, Michael J Wolyniak, Gregory Frechette, Christian R Lehman, Brandon K Fox, Paula Sundstrom
The mechanisms that fungi use to co-regulate subsets of genes specifically associated with morphogenic states represent a basic unsolved problem in fungal biology. Candida albicans is an important model of fungal differentiation both for rapid interconversion between yeast and hyphal growth forms and for white/opaque switching mechanisms. The Sundstrom lab is interested in mechanisms regulating hypha-specific expression of adhesin genes that are critical for C. albicans hyphal growth phenotypes and pathogenicity...
2018: PloS One
https://www.readbyqxmd.com/read/29434326/culture-independent-discovery-of-the-malacidins-as-calcium-dependent-antibiotics-with-activity-against-multidrug-resistant-gram-positive-pathogens
#7
Bradley M Hover, Seong-Hwan Kim, Micah Katz, Zachary Charlop-Powers, Jeremy G Owen, Melinda A Ternei, Jeffrey Maniko, Andreia B Estrela, Henrik Molina, Steven Park, David S Perlin, Sean F Brady
Despite the wide availability of antibiotics, infectious diseases remain a leading cause of death worldwide1 . In the absence of new therapies, mortality rates due to untreatable infections are predicted to rise more than tenfold by 2050. Natural products (NPs) made by cultured bacteria have been a major source of clinically useful antibiotics. In spite of decades of productivity, the use of bacteria in the search for new antibiotics was largely abandoned due to high rediscovery rates2,3 . As only a fraction of bacterial diversity is regularly cultivated in the laboratory and just a fraction of the chemistries encoded by cultured bacteria are detected in fermentation experiments, most bacterial NPs remain hidden in the global microbiome...
February 12, 2018: Nature Microbiology
https://www.readbyqxmd.com/read/29380150/dynamic-metabolic-control-towards-precision-engineering-of-metabolism
#8
Di Liu, Ahmad A Mannan, Yichao Han, Diego A Oyarzún, Fuzhong Zhang
Advances in metabolic engineering have led to the synthesis of a wide variety of valuable chemicals in microorganisms. The key to commercializing these processes is the improvement of titer, productivity, yield, and robustness. Traditional approaches to enhancing production use the "push-pull-block" strategy that modulates enzyme expression under static control. However, strains are often optimized for specific laboratory set-up and are sensitive to environmental fluctuations. Exposure to sub-optimal growth conditions during large-scale fermentation often reduces their production capacity...
January 29, 2018: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/29371619/immunization-with-transgenic-rodent-malaria-parasites-expressing-pfs25-induces-potent-transmission-blocking-activity
#9
K A Sala, F Angrisano, D F Da, I J Taylor, T S Churcher, A M Blagborough
An anti-malarial transmission blocking vaccine (TBV) would be an important tool for disease control or elimination, though current candidates have failed to induce high efficacy in clinical studies. The ookinete surface protein P25 is a primary target for TBV development, but heterologous expression of P25 with appropriate conformation is problematic and a pre-requisite for achieving functional titers. A potential alternative to recombinant/sub-unit vaccine is immunization with a non-pathogenic, whole-parasite vaccine...
January 25, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29286407/generic-protocol-for-optimization-of-heterologous-protein-production-using-automated-microbioreactor-technology
#10
Johannes Hemmerich, Lars Freier, Wolfgang Wiechert, Eric von Lieres, Marco Oldiges
A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development...
December 15, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29198849/oxalate-degrading-enzyme-recombined-lactic-acid-bacteria-strains-reduce-hyperoxaluria
#11
Chenming Zhao, Huan Yang, Xiaojing Zhu, Yang Li, Ning Wang, Shanfu Han, Hua Xu, Zhiqiang Chen, Zhangqun Ye
OBJECTIVE: To develop recombinant lactic acid bacteria (LAB) strains that express oxalate-degrading enzymes through biotechnology-based approach for the treatment of hyperoxaluria by oral administration. MATERIAL AND METHODS: The coding gene of oxalate decarboxylase (ODC) and oxalate oxidase (OxO) was transformed into Lactococcus lactis MG1363. The oxalate degradation ability in vitro was evaluated in media with high concentration of oxalate. Hyperoxaluria rat models through high oxalate diet were given recombinant LAB through oral administration...
December 2, 2017: Urology
https://www.readbyqxmd.com/read/29090064/-lactococcus-lactis-a-new-strategy-for-vaccination
#12
REVIEW
Maryam Azizpour, Seyyed Davood Hosseini, Parvaneh Jafari, Neda Akbary
Needle free vaccines have a several advantages and very attractive way for vaccination. In a body, mucosal surfaces provide a universal entry portal for all the known and emerging infectious pathogenic microbes. Therefore, it seems, vaccination strategies need to be reorganized for vaccines that are hindering the entry capability of pathogenic microbes through mucosal surfaces. Lactic acid Bacteria (LAB) are widely used in the food industry and at the present, used as delivery vehicles for biological investigations...
October 2017: Avicenna Journal of Medical Biotechnology
https://www.readbyqxmd.com/read/29040522/behavioral-and-transcriptomic-analysis-of-trem2-null-mice-not-all-knockout-mice-are-created-equal
#13
Silvia S Kang, Aishe Kurti, Kelsey E Baker, Chia-Chen Liu, Marco Colonna, Jason D Ulrich, David M Holtzman, Guojun Bu, John D Fryer
It is clear that innate immune system status is altered in numerous neurodegenerative diseases. Human genetic studies have demonstrated that triggering receptor expressed in myeloid cells 2 (TREM2) coding variants have a strong association with Alzheimer's disease (AD) and other neurodegenerative diseases. To more thoroughly understand the impact of TREM2 in vivo, we studied the behavioral and cognitive functions of wild-type (WT) and Trem2-/- (KO) mice during basal conditions and brain function in the context of innate immune stimulation with peripherally administered lipopolysaccharide (LPS)...
January 15, 2018: Human Molecular Genetics
https://www.readbyqxmd.com/read/29030449/plantaricyclin-a-a-novel-circular-bacteriocin-produced-by-lactobacillus-plantarum-ni326-purification-characterization-and-heterologous-production
#14
Juan Borrero, Eoin Kelly, Paula M O'Connor, Philip Kelleher, Colm Scully, Paul D Cotter, Jennifer Mahony, Douwe van Sinderen
Bacteriocins from lactic acid bacteria (LAB) are of increasing interest in recent years due to their potential as natural preservatives against food and beverage spoilage microorganisms. In a screening study for LAB, we isolated from olives a strain, Lactobacillus plantarum NI326, with activity against the beverage-spoilage bacterium Alicyclobacillus acidoterrestris Genome sequencing of NI326 enabled the identification of a gene cluster (designated plc ) encoding a putative circular bacteriocin and proteins involved in its modification, transport, and immunity...
January 1, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29021826/co-production-of-11%C3%AE-hydroxyprogesterone-and-ethanol-using-recombinant-yeast-expressing-fungal-steroid-hydroxylases
#15
Claire M Hull, Andrew G S Warrilow, Nicola J Rolley, Claire L Price, Iain S Donnison, Diane E Kelly, Steven L Kelly
BACKGROUND: Bioethanol production from sustainable sources of biomass that limit effect on food production are needed and in a biorefinery approach co-products are desirable, obtained from both the plant material and from the microbial biomass. Fungal biotransformation of steroids was among the first industrial biotransformations allowing corticosteroid production. In this work, the potential of yeast to produce intermediates needed in corticosteroid production is demonstrated at laboratory scale following bioethanol production from perennial ryegrass juice...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/29018422/novel-aggregation-promoting-factor-agge-contributes-to-the-probiotic-properties-of-enterococcus-faecium-bggo9-28
#16
Katarina Veljović, Nikola Popović, Marija Miljković, Maja Tolinački, Amarela Terzić-Vidojević, Milan Kojić
The understanding of mechanisms of interactions between various bacterial cell surface proteins and host receptors has become imperative for the study of the health promoting features of probiotic enterococci. This study, for the first time, describes a novel enterococcal aggregation protein, AggE, from Enterococcus faecium BGGO9-28, selected from a laboratory collection of enterococcal isolates with auto-aggregation phenotypes. Among them, En. faecium BGGO9-28 showed the strongest auto-aggregation, adhesion to components of ECM and biofilm formation...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28931397/pseudomonas-stutzeri-as-an-alternative-host-for-membrane-proteins
#17
Manuel Sommer, Hao Xie, Hartmut Michel
BACKGROUND: Studies on membrane proteins are often hampered by insufficient yields of the protein of interest. Several prokaryotic hosts have been tested for their applicability as production platform but still Escherichia coli by far is the one most commonly used. Nevertheless, it has been demonstrated that in some cases hosts other than E. coli are more appropriate for certain target proteins. RESULTS: Here we have developed an expression system for the heterologous production of membrane proteins using a single plasmid-based approach...
September 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28879631/biodegradation-of-phenol-and-its-derivatives-by-engineered-bacteria-current-knowledge-and-perspectives
#18
REVIEW
Lenka Rucká, Jan Nešvera, Miroslav Pátek
Biodegradation of phenolic compounds is a promising alternative to physical and chemical methods used to remove these toxic pollutants from the environment. The ability of various microorganisms to metabolize phenol and its derivatives (alkylphenols, nitrophenols and halogenated derivatives) has therefore been intensively studied. Knowledge of the enzymes catalyzing the individual reactions, the genes encoding these enzymes and the regulatory mechanisms involved in the expression of the respective genes in bacteria serves as a basis for the development of more efficient degraders of phenols via genetic engineering methods...
September 6, 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28874475/broad-host-range-expression-reveals-native-and-host-regulatory-elements-that-influence-heterologous-antibiotic-production-in-gram-negative-bacteria
#19
Jia Jia Zhang, Xiaoyu Tang, Michelle Zhang, Darlene Nguyen, Bradley S Moore
Heterologous expression has become a powerful tool for studying microbial biosynthetic gene clusters (BGCs). Here, we extend the transformation-associated recombination cloning and heterologous expression platform for microbial BGCs to include Gram-negative proteobacterial expression hosts. Using a broad-host-range expression platform, we test the implicit assumption that biosynthetic pathways are more successfully expressed in more closely related heterologous hosts. Cloning and expression of the violacein BGC from Pseudoalteromonas luteoviolacea 2ta16 revealed robust production in two proteobacterial hosts, Pseudomonas putida KT2440 and Agrobacterium tumefaciens LBA4404, but very little production of the antibiotic in various laboratory strains of Escherichia coli, despite their closer phylogenetic relationship...
September 5, 2017: MBio
https://www.readbyqxmd.com/read/28871183/functional-expression-of-the-entire-adhesiome-of-salmonella-enterica-serotype-typhimurium
#20
Nicole Hansmeier, Katarzyna Miskiewicz, Laura Elpers, Viktoria Liss, Michael Hensel, Torsten Sterzenbach
Adhesins are crucial virulence factors of pathogenic bacteria involved in colonization, transmission and pathogenesis. Many bacterial genomes contain the information for a surprisingly large number of diverse adhesive structures. One prominent example is the invasive and facultative intracellular pathogen Salmonella enterica with an adhesiome of up to 20 adhesins. Such large repertoire of adhesins contributes to colonization of a broad range of host species and may allow adaptation to various environments within the host, as well as in non-host environments...
September 4, 2017: Scientific Reports
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