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Recombinant baculovirus

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https://www.readbyqxmd.com/read/28922413/exploitation-of-stable-nanostructures-based-on-the-mouse-polyomavirus-for-development-of-a-recombinant-vaccine-against-porcine-circovirus-2
#1
Martin Fraiberk, Michaela Hájková, Magdaléna Krulová, Martina Kojzarová, Alena Drda Morávková, Ivan Pšikal, Jitka Forstová
The aim of this study was to develop a suitable vaccine antigen against porcine circovirus 2 (PCV2), the causative agent of post-weaning multi-systemic wasting syndrome, which causes significant economic losses in swine breeding. Chimeric antigens containing PCV2b Cap protein sequences based on the mouse polyomavirus (MPyV) nanostructures were developed. First, universal vectors for baculovirus-directed production of chimeric MPyV VLPs or pentamers of the major capsid protein, VP1, were designed for their exploitation as vaccines against other pathogens...
2017: PloS One
https://www.readbyqxmd.com/read/28917645/anti-fibrinolytic-and-anti-microbial-activities-of-a-serine-protease-inhibitor-from-honeybee-apis-cerana-venom
#2
Jie Yang, Kwang Sik Lee, Bo Yeon Kim, Yong Soo Choi, Hyung Joo Yoon, Jingming Jia, Byung Rae Jin
Bee venom contains a variety of peptide constituents, including low-molecular-weight protease inhibitors. While the putative low-molecular-weight serine protease inhibitor Api m 6 containing a trypsin inhibitor-like cysteine-rich domain was identified from honeybee (Apis mellifera) venom, no anti-fibrinolytic or anti-microbial roles for this inhibitor have been elucidated. In this study, we identified an Asiatic honeybee (A. cerana) venom serine protease inhibitor (AcVSPI) that was shown to act as a microbial serine protease inhibitor and plasmin inhibitor...
September 13, 2017: Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP
https://www.readbyqxmd.com/read/28904203/three-conserved-regions-in-baculovirus-sulfhydryl-oxidase-p33-are-critical-for-enzymatic-activity-and-function
#3
Wenhua Kuang, Huanyu Zhang, Manli Wang, Ning-Yi Zhou, Fei Deng, Hualin Wang, Peng Gong, Zhihong Hu
Baculoviruses encode a conserved sulfhydryl oxidase P33, which is necessary for budded virus (BV) production and multi-nucleocapsid occlusion-derived virus (ODV) formation. Here, the structural and functional relationship of P33 was revealed by X-ray crystallography, site-directed mutagenesis, and functional analysis. Based on crystallographic characterization and structural analysis, a series of P33 mutants within three conserved regions, i.e. the active site, dimer interface, and the R127-E183 salt bridge, were constructed...
September 13, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28890324/direct-head-to-head-evaluation-of-recombinant-adeno-associated-viral-vectors-manufactured-in-human-versus-insect-cells
#4
Oleksandr Kondratov, Damien Marsic, Sean M Crosson, Hector R Mendez-Gomez, Oleksandr Moskalenko, Mario Mietzsch, Regine Heilbronn, Jonathan R Allison, Kari B Green, Mavis Agbandje-McKenna, Sergei Zolotukhin
The major drawback of the Baculovirus/Sf9 system for recombinant adeno-associated viral (rAAV) manufacturing is that most of the Bac-derived rAAV vector serotypes, with few exceptions, demonstrate altered capsid compositions and lower biological potencies. Here, we describe a new insect cell-based production platform utilizing attenuated Kozak sequence and a leaky ribosome scanning to achieve a serotype-specific modulation of AAV capsid proteins stoichiometry. By way of example, rAAV5 and rAAV9 were produced and comprehensively characterized side by side with HEK293-derived vectors...
August 10, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28889175/expression-of-the-lef5-gene-from-spodoptera-exigua-multiple-nucleopolyhedrovirus-contributes-to-the-baculovirus-stability-in-cell-culture
#5
María Martínez-Solís, Agata K Jakubowska, Salvador Herrero
Baculoviruses are a broad group of viruses infecting insects, predominately of the order Lepidoptera. They are used worldwide as biological insecticides and as expression vectors to produce recombinant proteins. Baculoviruses replicate in their host, although several cell lines have been developed for in vitro replication. Nevertheless, replication of baculoviruses in cell culture involves the generation of defective viruses with a decrease in productivity and virulence. Transcriptional studies of the Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) and the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infective process revealed differences in the expression patterns when the virus replicated under in vitro (Se301 cells) or in vivo (S...
September 9, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28857510/cell-lines-from-diamondback-moth-exhibiting-differential-susceptibility-to-baculovirus-infection-and-expressing-midgut-genes
#6
Xiao-Li Ma, Wei-Yi He, Ping Wang, Min-Sheng You
Six new cell lines were established from embryonic tissues of the diamondback moth, Plutella xylostella (L.). The cell lines showed differential characteristics, including growth in attachment or in suspension, susceptibility to a baculovirus infection and expression of genes involved in the glucosinolate detoxication pathway in P. xylostella larvae. Five of the cells lines grew attached to the culture flask and one cell line grew unattached as a suspension cell line. The cell lines had population doubling times ranging from 18 to 23 h...
August 31, 2017: Insect Science
https://www.readbyqxmd.com/read/28843813/live-baculovirus-acts-as-a-strong-b-and-t-cell-adjuvant-for-monomeric-and-oligomeric-protein-antigens
#7
Suvi Heinimäki, Kirsi Tamminen, Maria Malm, Timo Vesikari, Vesna Blazevic
Recombinant proteins produced by baculovirus (BV) expression systems contain residual BV after crude purification. We studied adjuvant effect of BV on antibody and T cell responses against two model antigens, monomeric ovalbumin (OVA) protein and oligomeric norovirus (NoV) virus-like particles (VLPs). BALB/c mice were immunized intradermally with OVA alone or OVA formulated with live or inactivated BV, and VLP formulations comprised of chromatographically purified NoV GII.4 VLPs alone or mixed with BV, or of crude purified VLPs containing BV impurities from expression system...
August 24, 2017: Virology
https://www.readbyqxmd.com/read/28838345/prediction-of-g-gene-epitopes-of-viral-hemorrhagic-septicemia-virus-and-eukaryotic-expression-of-major-antigen-determinant-sequence
#8
T Sun, W-L Yin, B-H Fang, Q Wang, C-Z Liang, Z-Q Yue
This study aims to express fish Viral hemorrhagic septicemia virus (VHSV) G main antigen domain by using Bac-to-bac expression system. Using bioinformatics tools, B cell epitope of VHSV G gene was predicted, and G main antigen domain was optimized. GM gene was inserted into pFastBac1 vector, then transferred recombinant plasmid into DH10Bac to get recombinant rBacmid-GM. Obtained shuttle plasmid rBacmid-GM was transfected into sf9 cells. GM expression was examined using by PCR and western-blot. Results indicated that G main antigen domain gene of VHSV was successfully cloned and sequenced which contains 1209 bp...
August 15, 2017: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/28833672/a-large-scale-expression-strategy-for-multimeric-extracellular-protein-complexes-using-drosophila-s2-cells-and-its-application-to-the-recombinant-expression-of-heterodimeric-ligand-binding-domains-of-taste-receptor
#9
Atsuko Yamashita, Eriko Nango, Yuji Ahikawa
Many of the extracellular proteins or extracellular domains of plasma membrane proteins exist or function as homo- or heteromeric multimer protein complexes. Successful recombinant production of such proteins is often achieved by co-expression of the components using eukaryotic cells via the secretory pathway. Here we report a strategy addressing large-scale expression of hetero-multimeric extracellular domains of plasma membrane proteins and its application to the extracellular domains of a taste receptor...
August 19, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28830745/baculovirus-induced-recombinant-protein-expression-in-human-mesenchymal-stromal-stem-cells-a-promoter-study
#10
Gundula Sprick, Tobias Weidner, Denise Salzig, Peter Czermak
Human mesenchymal stem cells (hMSCs) are the current workhorses of regenerative medicine and gene therapy. The corresponding vectors are usually based on lentiviruses, adenoviruses, retroviruses or adeno-associated viruses, but recently they have been joined by baculoviruses, which are more widely known for their role in the development of pesticides and vaccines. Here we show that gene transfer to an immortalized human mesenchymal stroma cell line can be achieved by baculovirus transduction. We also compared the performance of five different constitutive promoters controlling GFP expression...
August 19, 2017: New Biotechnology
https://www.readbyqxmd.com/read/28823035/armored-dna-in-recombinant-baculoviruses-as-controls-in-molecular-genetic-assays
#11
Andrea Freystetter, Christian Paar, Herbert Stekel, Jörg Berg
The widespread use of molecular PCR-based assays in analytical and clinical laboratories brings about the need for test-specific, stable, and reliable external controls (EC) as well as standards and internal amplification controls (IC), in order to arrive at consistent test results. In addition, there is also a growing need to produce and provide stable, well-characterized molecular controls for quality assurance programs. In this study, we describe a novel approach to generate armored double-stranded DNA controls, which are encapsulated in baculovirus (BV) particles of the species Autographa californica multiple nucleopolyhedrovirus...
August 19, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28822988/the-components-of-shear-stress-affecting-insect-cells-used-with-the-baculovirus-expression-vector-system
#12
Tobias Weidner, Damir Druzinec, Martina Mühlmann, Rainer Buchholz, Peter Czermak
Insect-based expression platforms such as the baculovirus expression vector system (BEVS) are widely used for the laboratory- and industrial-scale production of recombinant proteins. Thereby, major drawbacks to gain high-quality proteins are the lytic infection cycle and the shear sensitivity of infected insect cells due to turbulence and aeration. Smaller bubbles were formerly assumed to be more harmful than larger ones, but we found that cell damage is also dependent on the concentration of protective agents such as Pluronic®...
September 26, 2017: Zeitschrift Für Naturforschung. C, A Journal of Biosciences
https://www.readbyqxmd.com/read/28814516/baculovirus-molecular-evolution-via-gene-turnover-and-recurrent-positive-selection-of-key-genes
#13
Tom Hill, Robert L Unckless
Hosts and viruses are locked in an evolutionary arms race. Hosts are constantly evolving to suppress virulence and replication, while viruses, reliant on host machinery for survival and reproduction, develop counter-strategies to escape this immune defense. Viruses must also adapt to novel conditions when establishing themselves in a host species. Both processes provide strong selection for viral adaptation. Understanding adaptive evolution in insect viruses can help us better understand adaptive evolution in general, and is important due to their use as biocontrol agents, and for protecting ecologically or economically important species from outbreaks...
August 16, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28802994/expression-and-purification-of-norovirus-virus-like-particles-in-escherichia-coli-and-their-immunogenicity-in-mice
#14
Yuqi Huo, Xin Wan, Tong Ling, Jie Wu, Wenhui Wang, Shuo Shen
Norovirus (NoV) virus like particles (VLPs) produced in Spodoptera frugiperda (Sf9) cells have been tested in human volunteers as vaccine candidate and were shown to be protective against NoV induced acute gastroenteritis. In this study, prevailing Sydney-2012-like NoV major capsid protein gene with or without N-terminal deletions (N26 and N38, 26 and 38 amino acids deleted from N terminus,respectively) were sub-cloned into prokaryotic expression vector, pCold III and pCold IV. Soluble and insoluble proteins were detected for both vectors after induction and higher levels of protein expression were observed for constructs pCold III-N26 and pCold III-N38...
August 9, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28794924/expression-of-tomato-yellow-leaf-curl-virus-coat-protein-using-baculovirus-expression-system-and-evaluation-of-its-utility-as-a-viral-antigen
#15
Lamiaa Elgaied, Reda Salem, Wael Elmenofy
DNA encoding the coat protein (CP) of an Egyptian isolate of tomato yellow leaf curl virus (TYLCV) was inserted into the genome of Autographa californica nucleopolyhedrovirus (AcNPV) under the control of polyhedrin promoter. The generated recombinant baculovirus construct harboring the coat protein gene was characterized using PCR analysis. The recombinant coat protein expressed in infected insect cells was used as a coating antigen in an indirect Enzyme-linked immunosorbent assay (ELISA) and dot blot to test its utility for the detection of antibody generated against TYLCV virus particles...
August 2017: 3 Biotech
https://www.readbyqxmd.com/read/28780630/binding-of-rabbit-hemorrhagic-disease-virus-like-particles-to-host-histo-blood-group-antigens-is-blocked-by-antisera-from-experimentally-vaccinated-rabbits
#16
Yanhua Song, Zhiyu Fan, Yuanyuan Zuo, Houjun Wei, Bo Hu, Mengmeng Chen, Rulong Qiu, Jiabin Xue, Fang Wang
During infection host histo-blood group antigens (HBGAs) act as attachment factors that interact with rabbit hemorrhagic disease virus (RHDV) and participate in the infectious process. In the present study, baculovirus expressing recombinant RHDV capsid protein (VP60r) as a vaccine immunogen was used to test its antigenicity and immunogenicity via immunization experiments. Each group of rabbits immunized with VP60r was found to be fully protected against RHDV challenge. The duration of immunity of the vaccine following the inoculation of a single dose was determined to be at least 240 days...
August 5, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28762492/purification-of-recombinant-human-tyrosinase-from-insect-larvae-infected-with-the-baculovirus-vector
#17
Monika B Dolinska, Paul T Wingfield, Yuri V Sergeev
The purification of an enzyme from insect larvae infected with a baculovirus vector is described. The enzyme tyrosinase is of biomedical importance and catalyzes the first rate-limiting steps in melanin production. Tyrosinase mutations can result in oculocutaneous albinism type 1 (OCA1), an inherited eye disease associated with decreased melanin pigment production and vision defects. To simplify expression and subsequent purification, the extracellular domain is expressed in insect cells, produced in Trichoplusia ni larvae, and purified using affinity and size-exclusion chromatography...
August 1, 2017: Current Protocols in Protein Science
https://www.readbyqxmd.com/read/28757575/vaccination-with-recombinant-baculovirus-expressing-ranavirus-major-capsid-protein-induces-protective-immunity-in-chinese-giant-salamander-andrias-davidianus
#18
Xiaoyuan Zhou, Xinglang Zhang, Yahui Han, Qiuhong Jia, Hongwei Gao
The Chinese giant salamander iridovirus (CGSIV), belonging to the genus Ranavirus in the family Iridoviridae, is the causative agent of an emerging infectious disease causing high mortality of more than 90% and economic losses in Chinese giant salamanders in China. In this study, a recombinant baculovirus-based vaccine expressing the CGSIV major capsid protein (MCP) was developed and its protective immunity in Chinese giant salamanders was evaluated. The recombinant Autographacalifornica nucleopolyhedrosis virus (AcNPV), expressing CGSIV MCP, designated as AcNPV-MCP, was generated with the highest titers of 1 × 10⁸ plaque forming units/mL (PFU/mL) and confirmed by Western blot and indirect immunofluorescence (IIF) assays...
July 25, 2017: Viruses
https://www.readbyqxmd.com/read/28755312/mouse-nudt13-is-a-mitochondrial-nudix-hydrolase-with-nad-p-h-pyrophosphohydrolase-activity
#19
Salama R Abdelraheim, David G Spiller, Alexander G McLennan
The mammalian NUDT13 protein possesses a sequence motif characteristic of the NADH pyrophosphohydrolase subfamily of Nudix hydrolases. Due to the persistent insolubility of the recombinant product expressed in Escherichia coli, active mouse Nudt13 was expressed in insect cells from a baculovirus vector as a histidine-tagged recombinant protein. In vitro, it efficiently hydrolysed NADH to NMNH and AMP and NADPH to NMNH and 2',5'-ADP and had a marked preference for the reduced pyridine nucleotides. Much lower activity was obtained with other nucleotide substrates tested...
July 28, 2017: Protein Journal
https://www.readbyqxmd.com/read/28753552/overexpression-of-c-myc-enhances-recombinant-protein-production-in-high-five-cells-after-baculovirus-infection
#20
Xiaoli Ju, Meijia Ren, Keping Chen, Qiang Wang
Due to their numerous advantages, baculovirus expression vector systems (BEVS) have been widely used to express recombinant proteins for different purposes. Different strategies have been adopted to increase recombinant protein production. In this study, we transiently or stably expressed mouse c-Myc in High Five cells using a commercial pIB/V5 vector. Under the control of the OpIE2 promoter, this vector could enhance recombinant protein production. We found that transient expression of c-Myc in High Five cells improved recombinant protein production...
July 28, 2017: Zeitschrift Für Naturforschung. C, A Journal of Biosciences
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