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Recombinant baculovirus

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https://www.readbyqxmd.com/read/28814516/baculovirus-molecular-evolution-via-gene-turnover-and-recurrent-positive-selection-of-key-genes
#1
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
#2
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
#3
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
#4
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
#5
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
#6
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
#7
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
#8
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
https://www.readbyqxmd.com/read/28685146/his-flag-tag-as-a-fusion-partner-of-glycosylated-human-interferon-gamma-and-its-mutant-gain-or-loss
#9
Elena Krachmarova, Milena Tileva, Elena Lilkova, Peicho Petkov, Klaus Maskos, Nevena Ilieva, Ivan Ivanov, Leandar Litov, Genoveva Nacheva
In order to obtain glycosylated human interferon-gamma (hIFNγ) and its highly prone to aggregation mutant K88Q, a secretory expression in insect cells was employed. To facilitate recombinant proteins purification, detection, and stability the baculovirus expression vectors were constructed to bear N-terminal His6-FLAG tag. Although the obtained proteins were glycosylated, we found that their biological activity was 100 times lower than expected. Our attempts to recover the biological properties of both proteins by tag removal failed due to enterokinase resistance of the tag...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28668116/macrobac-new-technologies-for-robust-and-efficient-large-scale-production-of-recombinant-multiprotein-complexes
#10
Scott D Gradia, Justin P Ishida, Miaw-Sheue Tsai, Chris Jeans, John A Tainer, Jill O Fuss
Recombinant expression of large, multiprotein complexes is essential and often rate limiting for determining structural, biophysical, and biochemical properties of DNA repair, replication, transcription, and other key cellular processes. Baculovirus-infected insect cell expression systems are especially well suited for producing large, human proteins recombinantly, and multigene baculovirus systems have facilitated studies of multiprotein complexes. In this chapter, we describe a multigene baculovirus system called MacroBac that uses a Biobricks-type assembly method based on restriction and ligation (Series 11) or ligation-independent cloning (Series 438)...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28664308/a-cell-clone-strain-from-mythimna-separata-lepidoptera-noctuidae-highly-susceptible-to-autographa-californica-multiple-nucleopolyhedrovirus-acmnpv-and-m-separata-npv-msnpv
#11
Xiang-Qian Meng, Gui-Ling Zheng, Chuan-De Zhao, Fang-Hao Wan, Chang-You Li
In this study, we describe a cell line, Ms-10C, cloned from the line QAU-Ms-E-10 (simplified Ms-10), an embryonic line from Mythimna separata. The cloned cell line was significantly more sensitive to nucleopolyhedrovirus (NPV). Ms-10C cells were mainly spherical with a diameter of 14.42 ± 2.23 μm. DNA amplification fingerprinting (DAF) confirmed the profile of PCR-amplified bands of the cloned cell line was consistent with those of the parental cell line, Ms-10. The sequencing result of the mitochondrial cytochrome c oxidase I (mtCO I) fragment confirmed that the amplified 636-bps mtCOI fragment was 100% identical to that of M...
August 2017: In Vitro Cellular & Developmental Biology. Animal
https://www.readbyqxmd.com/read/28660316/expression-of-flp-protein-in-a-baculovirus-insect-cell-system-for-biotechnological-applications
#12
Ida S Jensen, Ken Inui, Srdja Drakulic, Sakthidasan Jayaprakash, Bjoern Sander, Monika M Golas
The Saccharomyces cerevisiae Flp protein is a site-specific recombinase that recognizes and binds to the Flp recognition target (FRT) site, a specific sequence comprised of at least two inverted repeats separated by a spacer. Binding of four monomers of Flp is required to mediate recombination between two FRT sites. Because of its site-specific cleavage characteristics, Flp has been established as a genome engineering tool. Amongst others, Flp is used to direct insertion of genes of interest into eukaryotic cells based on single and double FRT sites...
August 2017: Protein Journal
https://www.readbyqxmd.com/read/28653811/biochemical-characterization-of-medaka-oryzias-latipes-orthologue-for-mammalian-factor-xiii-and-establishment-of-gene-edited-mutant
#13
Rima Horimizu, Ryota Ogawa, Yuko Watanabe, Hideki Tatsukawa, Masato Kinoshita, Hisashi Hashimoto, Kiyotaka Hitomi
At the final process of blood coagulation, fibrin molecules are stabilized via a catalytic reaction by Factor XIIIA (FXIIIA), one of the family members of transglutaminase (TGase) that catalyzes protein cross-linking reactions. In this study, we characterized the orthologue of this enzyme in medaka (Oryzias latipes), an established model fish in which a coagulation system is also preserved. The recombinant protein of this orthologue enzyme was produced in baculovirus-infected insect cells and used for biochemical property analysis including activation by thrombin-proteolysis and calcium-dependence of the TGase enzymatic activity...
June 27, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28649776/improving-the-baculovirus-expression-vector-system-with-vankyrin-enhanced-technology
#14
Kendra H Steele, Barbara J Stone, Kathleen M Franklin, Xiufeng Zhang, Haobo Jiang, Bruce A Webb, Christoph Geisler, Angelika Fath-Goodin
The baculovirus expression vector system (BEVS) is a widely used platform for the production of recombinant eukaryotic proteins. However, the BEVS has limitations in comparison to other higher eukaryotic expression systems. First, the insect cell lines used in the BEVS cannot produce glycoproteins with complex-type N-glycosylation patterns. Second, protein production is limited as cells die and lyse in response to baculovirus infection. To delay cell death and lysis, we transformed several insect cell lines with an expression plasmid harboring a vankyrin gene (P-vank-1), which encodes an anti-apoptotic protein...
June 26, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28630398/hcf-1-encoded-by-baculovirus-acmnpv-is-required-for-productive-nucleopolyhedrovirus-infection-of-non-permissive-tn368-cells
#15
Ami Tachibana, Rina Hamajima, Moe Tomizaki, Takuya Kondo, Yoshie Nanba, Michihiro Kobayashi, Hayato Yamada, Motoko Ikeda
Baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) replicates in both Spodoptera frugiperda Sf21 and Trichoplusia ni Tn368 cells, whereas AcMNPV defective in hcf-1 (host cell-factor 1) gene productively infects only Sf21 cells, indicating that HCF-1 is indispensable for the AcMNPV productive infection of Tn368 cells. Here, we demonstrated that HCF-1 protein transiently expressed in Tn368 cells promotes the DNA synthesis of Hyphantria cunea MNPV (HycuMNPV), Orygia pseudotsugata MNPV and Bombyx mori NPV, which are normally unable to replicate in Tn368 cells...
June 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28629377/quantitative-phosphoproteome-on-the-silkworm-bombyx-mori-cells-infected-with-baculovirus
#16
Jauharotus Shobahah, Shengjie Xue, Dongbing Hu, Cui Zhao, Ming Wei, Yanping Quan, Wei Yu
BACKGROUND: Bombyx mori has become an important model organism for many fundamental studies. Bombyx mori nucleopolyhedrovirus (BmNPV) is a significant pathogen to Bombyx mori, yet also an efficient vector for recombinant protein production. A previous study indicated that acetylation plays many vital roles in several cellular processes of Bombyx mori while global phosphorylation pattern upon BmNPV infection remains elusive. METHOD: Employing tandem mass tag (TMT) labeling and phosphorylation affinity enrichment followed by high-resolution LC-MS/MS analysis and intensive bioinformatics analysis, the quantitative phosphoproteome in Bombyx mori cells infected by BmNPV at 24 hpi with an MOI of 10 was extensively examined...
June 19, 2017: Virology Journal
https://www.readbyqxmd.com/read/28619136/coimmunization-with-recombinant-epitope-expressing-baculovirus-enhances-protective-effects-of-inactivated-h5n1-vaccine-against-heterologous-virus
#17
Zhen Zhang, Jiao Zhang, Jie Zhang, Qi Li, Peisi Miao, Jie Liu, Shijie Li, Jianni Huang, Ming Liao, Huiying Fan
H5N1, a highly pathogenic avian influenza virus (HPAIV), poses a significant threat to poultry and human health. However, currently available inactivated influenza vaccines are less efficacious against viruses that display antigenic drift. In this study, we constructed a recombinant baculovirus (BV-HMNN) expressing four conserved antigen epitopes: H5N1 hemagglutinin stem area amino acids 76-130 (HA2 76-130); three tandem repeats from the ectodomain of the conserved influenza matrix protein M2 (3M2e); nucleoprotein amino acids 55-69 (NP55-69); and nucleoprotein amino acids 380-393 (NP380-393)...
May 2017: Veterinary Microbiology
https://www.readbyqxmd.com/read/28605956/generation-of-pcv2-in-pk15-cells-transfected-with-recombinant-baculovirus-containing-a-1-1-copy-of-the-pcv2-genome
#18
Jie Cai, Xiaohong Xie, Yi Hu, Yang Zhan, Wanting Yu, Aibing Wang, Naidong Wang
Porcine circovirus associated diseases (PCVAD) caused by PCV2 are responsible for severe economic losses in the swine industry. The mechanism of PCV2 replication has not been fully elucidated yet. PCV2 may be successfully rescued by means of either an infectious DNA clone containing the full length of the viral genomic DNA, or from PCV2-infected clinical tissues in PK15 cell culture. However, viruses harvested by both methods have low titres. In this study, PCV2 was prepared with a higher titre from PK15 cells infected by recombinant baculoviruses containing 1PCV2 (one stem-loop structure) or 1...
June 2017: Acta Veterinaria Hungarica
https://www.readbyqxmd.com/read/28603894/a-performance-evaluation-of-a-novel-human-recombinant-tissue-factor-prothrombin-time-reagent-revohem-%C3%A2-pt
#19
C Gardiner, K Kohama, I Patel, P Lane, S Dwyer, S J Machin, I J Mackie
INTRODUCTION: A new prothrombin time reagent (Revohem™ PT) based on recombinant human tissue factor produced by the silkworm-baculovirus expression system was tested. The aim of this study was to compare the performance of the new PT reagent with two widely used routine PT reagents. METHODS: All testing was performed on a Sysmex CS-5100 coagulometer. Revohem(™) PT was tested for imprecision and stability using normal and abnormal lyophilized commercial control plasmas...
June 12, 2017: International Journal of Laboratory Hematology
https://www.readbyqxmd.com/read/28586369/synthesis-and-biochemical-characterization-of-egf-receptor-in-a-water-soluble-membrane-model-system
#20
Tiffany M Scharadin, Wei He, Yianni Yiannakou, Alexey A Tomilov, Matthew Saldana, Gino A Cortopassi, Kermit L Carraway, Matthew A Coleman, Paul T Henderson
ErbB (Erythroblastic Leukemia Viral Oncogene Homolog) receptor tyrosine kinases are critical for tissue development and maintenance, and frequently become oncogenic when mutated or overexpressed. In vitro analysis of ErbB receptor kinases can be difficult because of their large size and poor water solubility. Here we report improved production and assembly of the correctly folded full-length EGF receptor (EGFR) into nanolipoprotein particles (NLPs). NLPs are ~10 nm in diameter discoidal cell membrane mimics composed of apolipoproteins surrounding a lipid bilayer...
2017: PloS One
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