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Recombinant baculovirus

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https://www.readbyqxmd.com/read/28339926/study-on-the-expression-of-human-lysozyme-in-oviduct-bioreactor-mediated-by-recombinant-avian-adeno-associated-virus1
#1
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
#2
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
#3
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
#4
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...
February 22, 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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
https://www.readbyqxmd.com/read/28125901/onebac-2-0-sf9-cell-lines-for-production-of-aav1-aav2-and-aav8-vectors-with-minimal-encapsidation-of-foreign-dna
#11
Mario Mietzsch, Henrik Hering, Eva-Maria Hammer, Mavis Agbandje-McKenna, Sergei Zolotukhin, Regine Heilbronn
Recombinant adeno-associated viral (rAAV) vectors for human gene therapy require efficient and economical production methods to keep pace with the rapidly increasing clinical demand. In addition, the manufacturing process must ensure high vector quality and biological safety. The OneBac system offers easily scalable rAAV vector production in insect Sf9-derived AAV rep/cap-expressing producer cell lines infected with a single baculovirus that carries the rAAV backbone. For most AAV serotypes high burst sizes per cell were achieved, combined with high infectivity rates...
February 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28119062/baculovirus-based-genome-editing-in-primary-cells
#12
Maysam Mansouri, Zahra Ehsaei, Verdon Taylor, Philipp Berger
Genome editing in eukaryotes became easier in the last years with the development of nucleases that induce double strand breaks in DNA at user-defined sites. CRISPR/Cas9-based genome editing is currently one of the most powerful strategies. In the easiest case, a nuclease (e.g. Cas9) and a target defining guide RNA (gRNA) are transferred into a target cell. Non-homologous end joining (NHEJ) repair of the DNA break following Cas9 cleavage can lead to inactivation of the target gene. Specific repair or insertion of DNA with Homology Directed Repair (HDR) needs the simultaneous delivery of a repair template...
January 22, 2017: Plasmid
https://www.readbyqxmd.com/read/28092034/expression-and-purification-of-sh2-domains-using-baculovirus-expression-system
#13
Mari Ogiue-Ikeda, Kazuya Machida
Recombinant proteins expressed in bacteria are sometimes insoluble, aggregated, and incorrectly folded. For those Src homology 2 (SH2) domains that are insoluble in bacteria, baculovirus-insect cell expression systems can be an alternative to produce soluble and functionally active proteins. We describe a protocol for cloning and purification of GST-tagged SH2 domains using the Bac-to-Bac baculovirus expression system.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28089141/stable-emulsion-se-alone-is-an-effective-adjuvant-for-a-recombinant-baculovirus-expressed-h5-influenza-vaccine-in-healthy-adults-a-phase-2-trial
#14
John J Treanor, Laurence Chu, Brandon Essink, Derek Muse, Hana M El Sahly, Ruvim Izikson, Karen L Goldenthal, Peter Patriarca, Lisa M Dunkle
BACKGROUND: Influenza A viruses of the H5 subtype have been identified as important targets for development of vaccines. Achievement of potentially protective antibody responses against pandemic strains has usually required the use of adjuvants. OBJECTIVES: We evaluated a candidate A/Indonesia/05/2005 (H5) vaccine generated by baculovirus expression of recombinant hemagglutinin (HA) protein with or without stable emulsion (SE) as an adjuvant. METHODS: Healthy subjects 18-49years old were randomized (1:1:1:1) to receive two doses of rHA at 7...
February 7, 2017: Vaccine
https://www.readbyqxmd.com/read/28065477/quantitative-analysis-of-the-yield-of-avian-h7-influenza-virus-haemagglutinin-protein-produced-in-silkworm-pupae-with-the-use-of-the-codon-optimized-dna-a-possible-oral-vaccine
#15
Kuniaki Nerome, Sayaka Matsuda, Kenichi Maegawa, Shigeo Sugita, Kazumichi Kuroda, Kazunori Kawasaki, Reiko Nerome
In this study, we aimed to quantitatively compare the increased production of three H7 influenza virus-like particle (VLP) haemagglutinin (HA) with the use of a codon-optimized single HA gene in silkworm pupae. Recombinant baculovirus (Korea H7-BmNPV) could produce 0.40 million HA units per pupa, corresponding to 1832μg protein. The yield of the HA produced in larva was estimated to be approximately 0.31 million HA units per larva, and there were no significant differences between the three HA proteins. We could establish efficient recovery system of HA production in larvae and pupae with the use of three cycles sonication methods...
February 1, 2017: Vaccine
https://www.readbyqxmd.com/read/28061848/multi-antigen-avian-influenza-a-h7n9-virus-like-particles-particulate-characterizations-and-immunogenicity-evaluation-in-murine-and-avian-models
#16
Che-Ming Jack Hu, Chu-Yang Chien, Ming-Tsan Liu, Zih-Syun Fang, Sui-Yuan Chang, Rong-Huay Juang, Shih-Chung Chang, Hui-Wen Chen
BACKGROUND: Human infection with avian influenza A virus (H7N9) was first reported in China in March 2013. Since then, hundreds of cases have been confirmed showing severe symptoms with a high mortality rate. The virus was transmitted from avian species to humans and has spread to many neighboring areas, raising serious concerns over its pandemic potential. Towards containing the disease, the goal of this study is to prepare a virus-like particle (VLP) that consists of hemagglutinin (HA), neuraminidase (NA) and matrix protein 1 (M1) derived from the human isolate A/Taiwan/S02076/2013(H7N9) for potential vaccine development...
January 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28042093/expression-and-antigenicity-of-recombinant-human-respiratory-syncytial-virus-glycoproteins-having-different-affinity-tags
#17
Han Saem Lee, A-Reum Kim, Kisoon Kim, Wan-Ji Lee, Sung Soon Kim, You-Jin Kim
Human respiratory syncytial virus (HRSV) is a main cause of lower respiratory tract infections in infants and the elderly. Glycoprotein (G) is major antigen on the viral surface, and plays a key role for virus entry. Therefore, purification of the glycoprotein of HRSV is critical for the development of HRSV vaccine and serological diagnosis. In this study, we report the design and characterization of glycoprotein engineered rationally to enhance the protein solubility and to facilitate efficient purification...
December 29, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/28039392/molecular-characterization-of-tomato-%C3%AE-1-3-4-fucosidase-a-member-of-glycosyl-hydrolase-family-29-involved-in-the-degradation-of-plant-complex-type-n-glycans
#18
Md Ziaur Rahman, Megumi Maeda, Satsuki Itano, Anowar Hossain, Takeshi Ishimizu, Yoshinobu Kimura
In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Le(a) epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc...
December 30, 2016: Journal of Biochemistry
https://www.readbyqxmd.com/read/28018964/structure-determination-of-a-human-virus-by-the-combination-of-cryo-em-and-x-ray-crystallography
#19
Zheng Liu, Tom S Y Guu, Jianhao Cao, Yinyin Li, Lingpeng Cheng, Yizhi Jane Tao, Jingqiang Zhang
Virus 3D atomic structures provide insight into our understanding of viral life cycles and the development of antiviral drugs. X-ray crystallography and cryo-EM have been used to determine the atomic structure of viruses. However, limited availability of biological samples, biosafety issues due to virus infection, and sometimes inherent characteristics of viruses, pose difficulties on combining both methods in determining viral structures. These have made solving the high resolution structure of some medically important viruses very challenging...
2016: Biophysics Reports
https://www.readbyqxmd.com/read/27995596/development-of-biological-tools-to-assess-the-role-of-tmprss4-and-identification-of-novel-tumor-types-with-high-expression-of-this-prometastatic-protein
#20
Maria Villalba, Lissett Lopez, Miriam Redrado, Tamara Ruiz, Arrate L de Aberasturi, Nuria de la Roja, David Garcia, Francisco Exposito, Carlos de Andrea, Emilio Alvarez-Fernandez, Luis Montuenga, Paloma Rueda, Maria Jose Rodriguez, Alfonso Calvo
Metastatic spread is responsible for the majority of cancer deaths and identification of metastasis-related therapeutic targets is compulsory. TMPRSS4 is a pro-metastatic druggable transmembrane type II serine protease whose expression has been associated with the development of several cancer types and poor prognosis. To study the role and expression of this protease in cancer, we have developed molecular tools (active recombinant proteins and a polyclonal antibody) that can be used for diagnostic purposes and for testing anti-TMPRSS4 drugs...
December 20, 2016: Histology and Histopathology
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