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Recombinant expression

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https://www.readbyqxmd.com/read/28225274/a-rational-design-of-a-selective-inhibitor-for-kv1-1-channels-prevalent-in-demyelinated-nerves-that-improves-their-impaired-axonal-conduction
#1
Ahmed Al-Sabi, Declan Daly, Patrick Hoefer, Gemma K Kinsella, Charles P E Metais, Mark Pickering, Caroline Herron, Seshu Kumar Kaza, Kieran J Nolan, J Oliver Dolly
K(+) channels containing Kv1.1 α subunits, which become prevalent at internodes in demyelinated axons, may underlie their dysfunctional conduction akin to muscle weakness in multiple sclerosis. Small inhibitors were sought with selectivity for the culpable hyper-polarising K(+) currents. Modelling of interactions with the extracellular pore in a Kv1.1 deduced-structure identified a diaryldi-(2-pyrrolyl)methane (DPM) as a suitable scaffold, with optimised alkyl ammonium side chains. The resultant synthesised candidate [2,2'-((5,5'(di-p-topyldiaryldi-(2-pyrrolyl)methane)bis(2,2'carbonyl)bis (azanediyl)) diethaneamine...
February 22, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28225068/relish2-mediates-bursicon-homodimer-induced-prophylactic-immunity-in-the-mosquito-aedes-aegypti
#2
Hongwei Zhang, Shengzhang Dong, Xi Chen, David Stanley, Brenda Beerntsen, Qili Feng, Qisheng Song
Bursicon is a neuropeptide hormone consisting of two cystine-knot proteins (burs α and burs β), responsible for cuticle tanning and other developmental processes in insects. Recent studies show that each bursicon subunit forms homodimers that induce prophylactic immunity in Drosophila melanogaster. Here, we investigated the hypothesis that bursicon homodimers act in prophylactic immunity in insects, and possibly arthropods, generally, using the mosquito, Aedes aegypti. We found that burs α and burs β are expressed in larvae, pupae and newly emerged adults...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28224252/heat-shock-protein-70-in-lung-and-kidney-of-specific-pathogen-free-chickens-is-a-receptor-associated-protein-that-interacts-with-the-binding-domain-of-the-spike-protein-of-infectious-bronchitis-virus
#3
ZhiKun Zhang, Xin Yang, PengWei Xu, Xuan Wu, Long Zhou, HongNing Wang
Avian infectious bronchitis virus (IBV) is a member of the family Coronaviridae. A binding domain that mediates the attachment of the virus to its receptor has been identified in the S1 protein of prototype IBV strain M41. In this study, we identified this binding domain in a different strain, as well as the cellular proteins that interact with it. First, we expressed the S1N proteins (residues 19-270) of M41 and another isolate, SCZJ3, and compared the binding capacities of recombinant S1N-M41 and S1N-SCZJ3 to host tissues...
February 21, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28224118/heparin-binding-hemagglutinin-of-mycobacterium-tuberculosis-is-an-inhibitor-of-autophagy
#4
Qing Zheng, Zhi Li, Shan Zhou, Qian Zhang, Lei Zhou, Xiaorui Fu, Liu Yang, Yueyun Ma, Xiaoke Hao
Airway epithelial cell is often the initial site of attack by pathogens, and cell death is commonly caused by internalization of Mycobacterium tuberculosis (Mtb). However, the mechanism of interaction between epithelial cells and Mtb is not well understood. In this study, we investigated the role of the heparin-binding hemagglutinin (HBHA) protein of Mtb in the function of epithelial cells. In particular, the autophagy of A549 cells was determined based on microtubule-associated protein 1 light chain 3 alpha (LC3) activity...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28224001/overexpression-of-prunus-mume-dehydrin-genes-in-tobacco-enhances-tolerance-to-cold-and-drought
#5
Fei Bao, Dongliang Du, Yang An, Weiru Yang, Jia Wang, Tangren Cheng, Qixiang Zhang
Dehydrins, known as group 2 or D-11 family late-embryogenesis-abundant (LEA) proteins, play important roles in plant growth and stress tolerance. Six dehydrin genes were previously identified from the genome of Prunus mume. In this study, five of them (PmLEA8, PmLEA10, PmLEA19, PmLEA20, and PmLEA29) were cloned from cold-resistant P. mume 'Beijingyudie'. Real-time RT-PCR analysis indicated that all these genes could be up-regulated by one or several treatments (ABA, SA, low temperature, high temperature, PEG, and NaCl treatments)...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28223693/single-cd28-stimulation-induces-stable-and-polyclonal-expansion-of-human-regulatory-t-cells
#6
Xuehui He, Ruben L Smeets, Esther van Rijssen, Annemieke M H Boots, Irma Joosten, Hans J P M Koenen
CD4+FOXP3+ Treg are essential for immune tolerance. Phase-1 clinical trials of Treg-therapy to treat graft-versus-host-disease reported safety and potential therapeutic efficacy. Treg-based trials have started in organ-transplant patients. However, efficient ex vivo expansion of a stable Treg population remains a challenge and exploring novel ways for Treg expansion is a pre-requisite for successful immunotherapy. Based on the recent finding that CD28-signaling is crucial for survival and proliferation of mouse Treg, we studied single-CD28 stimulation of human Treg, without T cell receptor stimulation...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28223458/mutation-of-the-n-terminal-region-of-chikungunya-virus-capsid-protein-implications-for-vaccine-design
#7
Adam Taylor, Xiang Liu, Ali Zaid, Lucas Y H Goh, Jody Hobson-Peters, Roy A Hall, Andres Merits, Suresh Mahalingam
Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encephalitic alphaviruses, nuclear translocation induces host cell transcriptional shutoff; however, the role of capsid protein nucleolar localization in arthritogenic alphaviruses remains unclear. Using recombinant enhanced green fluorescent protein (EGFP)-tagged expression constructs and CHIKV infectious clones, we describe a nucleolar localization sequence (NoLS) in the N-terminal region of capsid protein, previously uncharacterized in CHIKV...
February 21, 2017: MBio
https://www.readbyqxmd.com/read/28223401/transcriptome-analyses-of-rhesus-monkey-pre-implantation-embryos-reveal-a-reduced-capacity-for-dna-double-strand-break-repair-in-primate-oocytes-and-early-embryos
#8
Xinyi Wang, Denghui Liu, Dajian He, Shengbo Suo, Xian Xia, Xiechao He, Jing-Dong Han, Ping Zheng
Pre-implantation embryogenesis encompasses several critical events including genome reprogramming, zygotic genome activation (ZGA), and cell fate commitment, most of which remain mechanistically unclear in primates. In addition, primates display a high rate of embryo wastage without any clear molecular basis. Understanding the factors involved in genome reprogramming and ZGA will help the generation of induced pluripotent stem cells with high efficiency. Moreover, explaining the molecular basis responsible for embryo wastage in primates will greatly expand our knowledge of species evolution...
February 21, 2017: Genome Research
https://www.readbyqxmd.com/read/28223313/active-multi-enzyme-assemblies-for-long-chain-olefinic-hydrocarbon-biosynthesis
#9
James K Christenson, Matthew R Jensen, Brandon R Goblirsch, Fatuma Mohamed, Wei Zhang, Carrie M Wilmot, Lawrence P Wackett
Bacteria from different Phyla produce long-chain olefinic hydrocarbons derived from an OleA-catalyzed Claisen condensation of two fatty acyl-CoA substrates, followed by reduction and oxygen elimination reactions catalyzed by the proteins OleB, OleC, and OleD. In this report, OleA, OleB, OleC, and OleD were individually purified as soluble proteins, and all were found to be essential for reconstituting hydrocarbon biosynthesis. Recombinant co-expression of tagged OleABCD proteins from Xanthomonas campestris in Escherichia coli and purification over His6x- and FLAG-columns resulted in OleA separating whilst OleBCD purified together, irrespective of which of the four Ole proteins were tagged...
February 21, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28223253/development-and-characterization-of-mouse-monoclonal-antibodies-reactive-with-chicken-cxcli2
#10
Woo H Kim, Hyun S Lillehoj, Yeasul Lim, Wongi Min, Yvonne B Sullivan, Laura Kakach, Joanna W LaBresh
Interleukin-8(IL-8)/CXCL8 is a CXC-family chemokine attracts lymphocytes to sites of tissue damage and plays a role in the inflammatory response and wound healing. Chicken chemotactic and angiogenic factor was referred to as chCXCLi2 and has been studied as one of human CXCL8 homologue for more than 20 years. However, no monoclonal antibodies (mAbs) that specifically detect chCXCLi2 have been developed. Here, we developed and characterized mouse mAbs against chCXCLi2 to define its immunological properties. Two mouse mAbs against chCXCLi2 were generated and confirmed to display specific binding with not only recombinants, but endogenous chCXCLi2 by Western blot analysis, ELISA, and immunocytochemistry...
February 18, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28223218/preventative-effect-of-omz-spt-on-lipopolysaccharide-induced-acute-lung-injury-and-inflammation-via-nuclear-factor-kappa-b-signaling-in-mice
#11
Ting Wang, Wanru Hou, Zhou Fu
Acute lung injury (ALI) is an early pathophysiologic change in acute respiratory distress syndrome and its management can be challenging. Omalizumab (Xolair™) is a recombinant DNA-derived, humanized antibody. OMZ-SPT is a polypeptide on the heavy chain of omalizumab monoclonal antibody. Here, we found that intramuscular administration of OMZ-SPT significantly improved survival and attenuated lung inflammation in female C57BL/6 mice suffering from lipopolysaccharide (LPS)-induced ALI. We also demonstrated that OMZ-SPT can inhibit expression of the inflammatory cytokines tumor necrosis factor-α, interleukin-1β and interleukin-6 by ELISA in mice suffering from LPS-induced ALI and a mouse macrophage line (RAW264...
February 18, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28222997/integrated-analysis-of-recombinant-bpv-1-l1-protein-for-the-production-of-a-bovine-papillomavirus-vlp-vaccine
#12
Diego Grando Módolo, Rodrigo Pinheiro Araldi, Jacqueline Mazzuchelli-de-Souza, Alexandre Pereira, Daniel Carvalho Pimenta, Letícia Maria Zanphorlin, Willy Beçak, Marcelo Menossi, Rita de Cassia Stocco, Rodrigo Franco de Carvalho
Bovine papillomatosis is an infectious disease that is caused by bovine papillomavirus (BPV), which results in important economic losses. However, no BPV vaccines or effective treatment methods are commercially available to date. Moreover, the absence of papillomavirus replication in vitro makes the use of recombinant protein a promising candidate for vaccine formulations. Hence, we developed an integrated study on the L1 capsid protein of BPV-1, obtained from a bacterial expression system, regarding its purification, biosafety, thermostability and immunogenicity...
February 17, 2017: Vaccine
https://www.readbyqxmd.com/read/28222316/discovery-and-characterization-of-novel-cyp1b1-inhibitors-based-on-heterocyclic-chalcones-overcoming-cisplatin-resistance-in-cyp1b1-overexpressing-lines
#13
Neill J Horley, Kenneth J M Beresford, Tarun Chawla, Glen J P McCann, Ketan C Ruparelia, Linda Gatchie, Vinay R Sonawane, Ibidapo S Williams, Hoon L Tan, Prashant Joshi, Sonali S Bharate, Vikas Kumar, Sandip B Bharate, Bhabatosh Chaudhuri
The structure of alpha-napthoflavone (ANF), a potent inhibitor of CYP1A1 and CYP1B1, mimics the structure of chalcones. Two potent CYP1B1 inhibitors 7k (DMU2105) and 6j (DMU2139) have been identified from two series of synthetic pyridylchalcones. They inhibit human CYP1B1 enzyme bound to yeast-derived microsomes (Sacchrosomes™) with IC50 values of 10 and 9 nM, respectively, and show a very high level of selectivity towards CYP1B1 with respect to the IC50 values obtained with CYP1A1, CYP1A2, CYP3A4, CYP2D6, CYP2C9 and CYP2C19 Sacchrosomes™...
February 9, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28221723/animal-component-free-agrobacterium-tumefaciens-cultivation-media-for-better-gmp-compliance-increases-biomass-yield-and-pharmaceutical-protein-expression-in-nicotiana-benthamiana
#14
Marcel Houdelet, Anna Galinski, Tanja Holland, Kathrin Wenzel, Stefan Schillberg, Johannes Felix Buyel
Transient expression systems allow the rapid production of recombinant proteins in plants. Such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. However, large-scale transient expression requires the production of recombinant Agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. The complex media often used for the cultivation of this species typically include animal-derived ingredients that can contain human pathogens, thus conflicting with the requirements of good manufacturing practice (GMP)...
February 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28220810/dengue-virus-ns1-protein-activates-immune-cells-via-tlr4-but-not-tlr2-or-tlr6
#15
Naphak Modhiran, Daniel Watterson, Antje Blumenthal, Alan G Baxter, Paul R Young, Katryn J Stacey
The secreted hexameric form of the dengue virus (DENV) non-structural protein 1 (NS1) has recently been shown to elicit inflammatory cytokine release and disrupt endothelial cell monolayer integrity. This suggests that circulating NS1 contributes to the vascular leak that plays a major role in the pathology of dengue haemorrhagic fever and shock. Pathways activated by NS1 are thus of great interest as potential therapeutic targets. Recent works have separately implicated both toll-like receptor 4 (TLR4) and the TLR2/6 heterodimer in immune cell activation by NS1...
February 21, 2017: Immunology and Cell Biology
https://www.readbyqxmd.com/read/28220242/expression-of-clonorchis-sinensis-giiispla2-protein-in-baculovirus-infected-insect-cells-and-its-overexpression-facilitating-epithelial-mesenchymal-transition-in-huh7-cells-via-akt-pathway
#16
Mei Shang, Zhizhi Xie, Zeli Tang, Lei He, Xiaoyun Wang, Caiqin Wang, Yinjuan Wu, Ye Li, Lu Zhao, Zhiyue Lv, Zhongdao Wu, Yan Huang, Xinbing Yu, Xuerong Li
Although prior studies confirmed that group III secretory phospholipase A2 of Clonorchis sinensis (CsGIIIsPLA2) had stimulating effect on liver fibrosis by binding to LX-2 cells, large-scale expression of recombinant protein and its function in the progression of hepatoma are worth exploring. Because of high productivity and low lipopolysaccharides (LPS) in the Sf9-baculovirus expression system, we firstly used this system to express the coding region of CsGIIIsPLA2. The molecular weight of recombinant CsGIIIsPLA2 protein was about 34 kDa...
February 20, 2017: Parasitology Research
https://www.readbyqxmd.com/read/28220142/absence-of-an-n-linked-glycosylation-motif-in-the-glycoprotein-of-the-live-attenuated-argentine-hemorrhagic-fever-vaccine-candid-1-results-in-its-improper-processing-and-reduced-surface-expression
#17
John T Manning, Alexey V Seregin, Nadezhda E Yun, Takaaki Koma, Cheng Huang, José Barral, Juan C de la Torre, Slobodan Paessler
Junin virus (JUNV), a highly pathogenic New World arenavirus, is the causative agent of Argentine hemorrhagic fever (AHF). The live-attenuated Candid #1 (Can) strain currently serves as a vaccine for at-risk populations. We have previously shown that the Can glycoprotein (GPC) gene is the primary gene responsible for attenuation in a guinea pig model of AHF. However, the mechanisms through which the GPC contributes to the attenuation of the Can strain remain unknown. A more complete understanding of the mechanisms underlying the attenuation and immunogenicity of the Can strain will potentially allow for the rational design of additional safe and novel vaccines...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28219398/interaction-between-hcmv-pul83-and-human-aim2-disrupts-the-activation-of-the-aim2-inflammasome
#18
Yuan Huang, Di Ma, Heyu Huang, Yuanyuan Lu, Yi Liao, Lingling Liu, Xinglou Liu, Feng Fang
BACKGROUND: AIM2, a cytosolic DNA sensor, plays an important role during infection caused by pathogens with double-stranded DNA; however, its role in human cytomegalovirus (HCMV) infection remains unclear. Previously, we showed an increase in AIM2 protein levels during the early stage of HCMV infection and a decrease 24 h post infection. Because HCMV has developed a variety of strategies to evade host immunity, we speculated that this decline might be attributed to a viral immune escape mechanism...
February 20, 2017: Virology Journal
https://www.readbyqxmd.com/read/28219382/high-level-extracellular-protein-production-in-bacillus-subtilis-using-an-optimized-dual-promoter-expression-system
#19
Kang Zhang, Lingqia Su, Xuguo Duan, Lina Liu, Jing Wu
BACKGROUND: We recently constructed a Bacillus subtilis strain (CCTCC M 2016536) from which we had deleted the srfC, spoIIAC, nprE, aprE and amyE genes. This strain is capable of robust recombinant protein production and amenable to high-cell-density fermentation. Because the promoter is among the factors that influence the production of target proteins, optimization of the initial promoter, PamyQ from Bacillus amyloliquefaciens, should improve protein expression using this strain. This study was undertaken to develop a new, high-level expression system in B...
February 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28219131/-recombinant-human-bone-morphogenetic-protein-7-inhibits-the-proliferation-and-extracellular-matrix-secretion-of-fibroblast-of-urethral-scars
#20
Y Wu, L M Ma, B Cai, H L Li, Q W Xing
Objective: To study the effect and its mechanism of recombinant human bone morphogenetic protein 7 (rhBMP-7) on urethral keloid fibroblast. Methods: Urethral fibroblasts were extracted and cultured, and the fourth-generation fibroblasts were used for experiment. Urethral fibroblasts were treated with rhBMP-7. Proliferation rate was detected by CCK-8 method. RT-PCR was applied to test changes of mRNA in α smooth muscle actin (α-SMA) and type Ⅰ collagen protein (COL-1) of each group. Western blot was used to measure α-SMA protein expression and the effect of rhBMP-7 on signaling pathway of urethral keloid fibroblasts...
February 14, 2017: Zhonghua Yi Xue za Zhi [Chinese medical journal]
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