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Recombinant baculovirus

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https://www.readbyqxmd.com/read/28432958/membrane-fusion-between-baculovirus-budded-virus-enveloped-particles-and-giant-liposomes-generated-using-a-droplet-transfer-method-for-the-incorporation-of-recombinant-membrane-proteins
#1
Misako Nishigami, Takaaki Mori, Masahiro Tomita, Kingo Takiguchi, Kanta Tsumoto
Giant proteoliposomes are generally useful as artificial cell membranes in biochemical and biophysical studies, and various procedures for their preparation have been reported. We present here a novel preparation technique that involves the combination of i) cell-sized lipid vesicles (giant unilamellar vesicles, GUVs) that are generated using the droplet-transfer method, where lipid monolayer-coated water-in-oil microemulsion droplets interact with oil/water interfaces to form enclosed bilayer vesicles, and ii) budded viruses (BVs) of baculovirus (Autographa californica nucleopolyhedrovirus) that express recombinant transmembrane proteins on their envelopes...
April 13, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28424279/phosphorylation-induces-structural-changes-in-the-autographa-californica-nucleopolyhedrovirus-p10-protein
#2
Farheen Raza, Joanna F McGouran, Benedikt M Kessler, Robert D Possee, Linda A King
Baculoviruses encode a variety of auxiliary proteins that are not essential for viral replication but provide them with a selective advantage in nature. P10 is a 10 kDa auxiliary protein produced in the very-late phase of gene transcription by Autographa californica multiple nucleopolyhedrovirus (AcMNPV). The P10 protein forms cytoskeletal-like structures in the host cell that associate with microtubules varying from filamentous forms in the cytoplasm to aggregated peri-nuclear tubules that form a cage-like structure around the nucleus...
April 19, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28412328/budded-baculovirus-particles-as-a-source-of-membrane-proteins-for-radioligand-binding-assay-the-case-of-dopamine-d1-receptor
#3
Anni Allikalt, Ago Rinken
INTRODUCTION: G-protein-coupled receptors have become very important drug targets and therefore ligand binding assays for these receptors are an essential part of drug discovery. Among a variety of experimental systems, the radioligand binding assay has remained as one of the main methods in this field for decades. Usually cell membranes or tissues are used in these experiments, however in this article we demonstrate that baculoviruses produced in Sf9 cells display recombinant receptors on their surface and therefore can be used in radioligand binding assay...
April 12, 2017: Journal of Pharmacological and Toxicological Methods
https://www.readbyqxmd.com/read/28393684/generation-of-the-fluorescent-hmgb1-gfp-fusion-protein-in-insect-cells-and-evaluation-of-its-immunogenicity-in-two-mice-models
#4
Ali Anvar, Rouhollah Vahabpour, Iman Salahshourifar, Azam Bolhassani
High mobility group box 1 (HMGB1) is a highly conserved protein present in the nuclei and cytoplasm of cells which has an important role as a mediator of inflammation in the extracellular environment. HMGB1 was identified as an innate adjuvant that induces immune responses against soluble antigens in vivo. In the current study, we used a baculovirus expression system for insect cells that was based on expression of HMGB1 with target gene (GFP), and purified the recombinant HMGB1-GFP fusion protein. We then demonstrated whether immunogenicity of GFP changes in the presence or absence of recombinant HMGB1 acting as an adjuvant in C57BL/6 and BALB/c mice...
April 4, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28385002/generation-characterization-and-application-in-serodiagnosis-of-recombinant-swine-vesicular-disease-viral-like-particles
#5
Wanhong Xu, Melissa Goolia, Tim Salo, Zhidong Zhang, Ming Yang
Swine vesicular disease virus (SVDV) is a highly contagious disease that causes vesicular disease in pigs. The importance of the disease is due to its indistinguishable clinical signs from foot-and-mouth disease that prevents international trade of swine and related products. SVD-specific antibody detection using the enzyme-linked immunosorbent assay (ELISA) is the most versatile and commonly used method for SVD surveillance and export certification. The inactivated SVD virus is the commonly used antigen in ELISAs...
April 6, 2017: Journal of Veterinary Science
https://www.readbyqxmd.com/read/28383905/construction-and-rescue-of-a-functional-synthetic-baculovirus
#6
Yu Shang, Manli Wang, Gengfu Xiao, Xi Wang, Dianhai Hou, Kai Pan, Shurui Liu, Jiang Li, Jun Wang, Basil M Arif, Just M Vlak, Xinwen Chen, Hualin Wang, Fei Deng, Zhihong Hu
Synthetic viruses provide a powerful platform to delve deeper into the nature and function of viruses as well as to engineer viruses with novel properties. So far, most synthetic viruses have been RNA viruses (<30 kb) and small DNA viruses, such as bacteriophage phiX174. Baculoviruses contain a large circular dsDNA genome of 80-180 kb and have been used as biocontrol agents and protein expression vectors. Here, we report on the first synthesis of a baculovirus based on the type species Autographa californica nucleopolyhedrovirus, AcMNPV, by a combination of PCR and transformation-associated recombination in yeast...
April 6, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28374513/identification-and-characterization-of-laodelphax-striatellus-insecta-hemiptera-delphacidae-neutral-sphingomyelinase
#7
Y Zhou, X-W Lin, M-A Begum, C-H Zhang, X-X Shi, W-J Jiao, Y-R Zhang, J-Q Yuan, H-Y Li, Q Yang, C Mao, Z-R Zhu
The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus, a direct sap-sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase-enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively...
April 4, 2017: Insect Molecular Biology
https://www.readbyqxmd.com/read/28352544/efficient-production-of-recombinant-pp2a-at-a-low-temperature-using-a-baculovirus-expression-system
#8
Tsuyoshi Ikehara, Shihoko Nakashima, Junichi Nakashima, Tsubasa Kinoshita, Takeshi Yasumoto
Protein phosphatase 2A (PP2A) is an enzyme useful for detecting several natural toxins represented by okadaic acid and microcystins. We found that the production of the recombinant human PP2A catalytic subunit (rhPP2Ac) in High Five insect cells could markedly increase when the cells were cultured at 19 °C instead of 27 °C used under conventional conditions. The yield and purity of the enzyme increased four- and three-folds, respectively. The benefit of the altered culturing temperature was observed with the recombinant human protein phosphatase 2B but not 2Cα...
September 2016: Biotechnology Reports
https://www.readbyqxmd.com/read/28345293/detection-of-zaire-ebolavirus-in-swine-assay-development-and-optimization
#9
B S Pickering, B Collignon, G Smith, P Marszal, G Kobinger, H M Weingartl
Ebolaviruses (family Filoviridae, order Mononegavirales) cause often fatal, haemorrhagic fever in primates including humans. Pigs have been identified as a species susceptible to Reston ebolavirus (RESTV) infection, with indicated transmission to humans in the Philippines; however, their role during Ebola outbreaks in Africa needs to be clarified. To perform surveillance studies, detection of ebolavirus requires a prerequisite validation of viral RNA and antibody detection methods in swine samples. These diagnostic tests also need to be suitable for deployment to low-level containment laboratories...
March 27, 2017: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/28342150/expression-and-characterization-of-human-%C3%AE-1-4-galactosyltransferase-1-%C3%AE-4galt1-using-silkworm-baculovirus-expression-system
#10
Daisuke Morokuma, Jian Xu, Masato Hino, Hiroaki Mon, Jasmeen S Merzaban, Masateru Takahashi, Takahiro Kusakabe, Jae Man Lee
Baculovirus expression vector system (BEVS) is widely known as a mass-production tool to produce functional recombinant glycoproteins except that it may not be always suitable for medical practice due to the differences in the structure of N-linked glycans between insects and mammalian. Currently, various approaches have been reported to alter N-linked glycan structures of glycoproteins derived from insects into terminally sialylated complex-type N-glycans. In the light of those studies, we also proposed in vitro maturation of N-glycan with mass-produced and purified glycosyltransferases by silkworm-BEVS...
March 24, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28339926/study-on-the-expression-of-human-lysozyme-in-oviduct-bioreactor-mediated-by-recombinant-avian-adeno-associated-virus1
#11
A P Wang, Y J Wang, S Wu, W Y Zuo, C M Guo, W M Hong, S Y Zhu
Due to its antimicrobial properties and low toxicity, human lysozyme (hLYZ) has broad application in the medical field and as a preservative used by the food industry. However, limited availability hinders its widespread use. Hence, we constructed a recombinant avian adeno-associated virus (rAAAV) that would specifically express hLYZ in the chicken oviduct and harvested hLYZ from the egg whites of laying hens. The oviduct-specific human lysozyme expression cassette flanked by avian adeno-associated virus (AAAV) inverted terminal repeats (ITRs) was subcloned into the modified baculovirus transfer vector pFBX, and then the recombinant baculovirus rBac-ITRLYZ was generated...
March 2, 2017: Poultry Science
https://www.readbyqxmd.com/read/28316015/expression-and-characterization-of-codon-optimized-crimean-congo-hemorrhagic-fever-virus-gn-glycoprotein-in-insect-cells
#12
Mehdi Rahpeyma, Alireza Samarbaf-Zadeh, Manoochehr Makvandi, Ata A Ghadiri, Stuart D Dowall, Fatemeh Fotouhi
Crimean-Congo hemorrhagic fever virus (CCHFV) is a major cause of tick-borne viral hemorrhagic disease in the world. Despite of its importance as a deadly pathogen, there is currently no licensed vaccine against CCHF disease. The attachment glycoprotein of CCHFV (Gn) is a potentially important target for protective antiviral immune responses. To characterize the expression of recombinant CCHFV Gn in an insect-cell-based system, we developed a gene expression system expressing the full-length coding sequence under a polyhedron promoter in Sf9 cells using recombinant baculovirus...
March 18, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28262785/a-novel-ideal-radionuclide-imaging-system-for-non-invasively-cell-monitoring-built-on-baculovirus-backbone-by-introducing-sleeping-beauty-transposon
#13
Jing Lv, Yu Pan, Huijun Ju, Jinxin Zhou, Dengfeng Cheng, Hongcheng Shi, Yifan Zhang
Sleeping Beauty (SB) transposon is an attractive tool in stable transgene integration both in vitro and in vivo; and we introduced SB transposon into recombinant sodium-iodide symporter baculovirus system (Bac-NIS system) to facilitate long-term expression of recombinant sodium-iodide symporter. In our study, two hybrid baculovirus systems (Bac-eGFP-SB-NeoR and Bac-NIS-SB-NeoR) were successfully constructed and used to infect U87 glioma cells. After G418 selection screening, the Bac-eGFP-SB-NeoR-U87 cells remained eGFP positive, at the 18(th) and 196(th) day post transfection (96...
March 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28260093/recombinant-adeno-associated-virus-serotype-9-in-a-mouse-model-of-atherosclerosis-determination-of-the-optimal-expression-time-in%C3%A2-vivo
#14
Qingjie Chen, Hui Zhai, Xiaomei Li, Yitong Ma, Bangdang Chen, Fen Liu, Hongmei Lai, Jia Xie, Chunhui He, Junyi Luo, Jing Gao, Yining Yang
Adeno-associated virus 9 (AAV9) has been identified as one of the optimal gene transduction carriers for gene therapy. The aim of the present study was to determine the gene transfection efficiency and safety of an AAV9 vector produced using a recombinant baculovirus (rBac)‑based system. AAV9‑cytomegalovirus (CMV)-green fluorescent protein was produced using an rBac system and the resulting vector particles were injected intravenously into mice. Animals were sacrificed at 14, 21, 28, 35, 60, 90 and 120 days following injection...
April 2017: Molecular Medicine Reports
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
#15
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/28216094/fusion-of-flagellin-2-with-bivalent-white-spot-syndrome-virus-vaccine-increases-survival-in-freshwater-shrimp
#16
Hansam Cho, Na Hye Park, Yuyeon Jang, Yong-Dae Gwon, Yeondong Cho, Yoon-Ki Heo, Ki-Hoon Park, Hee-Jung Lee, Tae Jin Choi, Young Bong Kim
Despite large economic losses attributable to white spot syndrome virus (WSSV), an infectious pathogen of penaeid shrimp and other crustaceans worldwide, no efficient vaccines or antiviral agents to control the virus are available at present. Here, we designed and constructed baculovirus-based vaccines delivering genes encoding the WSSV envelope proteins, VP28 and VP19. To enhance the immunogenicity of the baculovirus-based vaccine, we fused a Salmonella typhimurium flagellin 2 (FL2) gene with VP28 or VP19 gene...
March 2017: Journal of Invertebrate Pathology
https://www.readbyqxmd.com/read/28216055/human-eosinophils-constitutively-express-a-unique-serine-protease-prss33
#17
Sumika Toyama, Naoko Okada, Akio Matsuda, Hideaki Morita, Hirohisa Saito, Takao Fujisawa, Susumu Nakae, Hajime Karasuyama, Kenji Matsumoto
BACKGROUND: Eosinophils play important roles in asthma, especially airway remodeling, by producing various granule proteins, chemical mediators, cytokines, chemokines and proteases. However, protease production by eosinophils is not fully understood. In the present study, we investigated the production of eosinophil-specific proteases/proteinases by transcriptome analysis. METHODS: Human eosinophils and other cells were purified from peripheral blood by density gradient sedimentation and negative/positive selections using immunomagnetic beads...
February 16, 2017: Allergology International: Official Journal of the Japanese Society of Allergology
https://www.readbyqxmd.com/read/28182872/enhanced-critical-size-calvarial-bone-healing-by-ascs-engineered-with-cre-loxp-based-hybrid-baculovirus
#18
Shih-Chun Lo, Kuei-Chang Li, Yu-Han Chang, Mu-Nung Hsu, Li-Yu Sung, Truong Anh Vu, Yu-Chen Hu
Calvarial bone repair remains challenging for adults. Although adipose-derived stem cells (ASCs) hold promise to heal bone defects, use of ASCs for critical-size calvarial bone repair is ineffective. Stromal cell-derived factor 1 (SDF-1) is a chemokine capable of triggering stem cell migration. Although recombinant SDF-1 protein is co-delivered with other molecules including BMP-2 to facilitate calvarial bone repair, these approaches did not yield satisfactory healing. This study aimed to exploit a newly developed Cre/loxP-based hybrid baculovirus for efficient gene delivery and prolonged transgene expression in ASCs...
January 30, 2017: Biomaterials
https://www.readbyqxmd.com/read/28164534/expression-of-hcv-alternative-reading-frame-protein-core-1-f-in-baculovirus-expression-system-and-its-evaluation-for-assessment-of-specific-anti-core-1-antibody-in-iranian-hcv-infected-patients
#19
Pooneh Rahimi, Rouhollah Vahabpour, Farideh Sadat Sajadian Fard, Fatemeh Motevalli, Fatemeh Fotouhi Chahouki, Mohammad Reza Aghasadeghi, Azam Bolhassani, Seyed Mehdi Sadat, Ehsan Mostafavi, Nasir Mohajel, Ali Jahanian-Najafabadi, Mohammad Reza Amiran
BACKGROUND: Hepatitis C virus (HCV) genome contains an overlapping reading frame which results in alternative core protein (ARFP). Baculovirus expression system was used as a powerful eukaryotic vector system to express core+1/F protein for the first time. This recombinant core+1/F protein was used to assess the anti-core+1 antibody in anti-HCV drug resistant and sustained virologic response (SVR) patients. METHODS: The core+1 coding sequence from HCV genotype 1 was designed and synthesized in pUC57 vector...
October 1, 2016: Clinical Laboratory
https://www.readbyqxmd.com/read/28138375/designing-construction-and-expression-of-a-recombinant-fusion-protein-comprising-the-hepatitis-e-virus-orf2-and-rotavirus-nsp4-in-the-baculovirus-expression-system
#20
Manoochehr Makvandi, Ali Teimoori, Niloofar Neisi, Alireza Samarbafzadeh
BACKGROUND: The hepatitis E virus (HEV) accounts for hepatitis E infection with relatively high mortality rate in pregnant women that can lead to fulminant hepatitis. The baculovirus expression system (BES) has the capability to produce high-level recombinant proteins and could be useful for vaccine designing. OBJECTIVES: The aim of this study was designing a recombinant hepatitis E virus ORF2 and Rotavirus NSP4 (ORF2-NSP4) and to evaluating construction these recombinant proteins in the BES...
November 2016: Jundishapur Journal of Microbiology
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