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Recombinant baculovirus

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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
#1
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...
January 5, 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
#2
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
#3
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
#4
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
#5
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
#6
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
https://www.readbyqxmd.com/read/27989837/prophenoloxidase-activation-and-antimicrobial-peptide-expression-induced-by-the-recombinant-microbe-binding-protein-of-manduca-sexta
#7
Yang Wang, Haobo Jiang
Manduca sexta microbe binding protein (MBP) is a member of the β-1,3-glucanase-related protein superfamily that includes Gram-negative bacteria-binding proteins (GNBPs), β-1,3-glucan recognition proteins (βGRPs), and β-1,3-glucanases. Our previous and current studies showed that the purified MBP from baculovirus-infected insect cells had stimulated prophenoloxidase (proPO) activation in the hemolymph of naïve and immune challenged larvae and that supplementation of the exogenous MBP and peptidoglycans (PGs) had caused synergistic increases in PO activity...
October 28, 2016: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27984103/generation-and-characterization-of-new-monoclonal-antibodies-against-swine-origin-2009-influenza-a-h1n1-virus-and-evaluation-of-their-prophylactic-and-therapeutic-efficacy-in-a-mouse-model
#8
Sheng-Fan Wang, Sung-Pin Tseng, El-Wui Loh, Wen-Hung Wang, Ming-Chun Li, Kuan-Hsuan Chen, Wan-Chi Tsai, Yuan-Ming Lee, Huan-Yuan Chen, Fu-Tong Liu, Yi-Ming Arthur Chen, Jason C Huang
In 2009, a swine-origin influenza A virus - A(H1N1)pdm09 - emerged and has became a pandemic strain circulating worldwide. The hemagglutinin (HA) of influenza virus is a potential target for the development of anti-viral therapeutic agents. Here, we generated mAbs by immunization of baculovirus-insect expressing trimeric recombinant HA of the A(H1N1)pdm09 strain. Results indicated that the mAbs recognized two novel neutralizing and protective epitopes-"STAS" and "FRSK" which located near Cb and Ca1 antigenic regions respectively and were conserved in almost 2009-2016 influenza H1N1 stains...
October 27, 2016: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/27974824/induction-of-a-robust-immunity-response-against-novel-duck-reovirus-in-ducklings-using-a-subunit-vaccine-of-sigma-c-protein
#9
Zhuangli Bi, Yingqi Zhu, Zongyan Chen, Chuanfeng Li, Yong Wang, Guijun Wang, Guangqing Liu
Novel duck reovirus (NDRV) disease emerged in China in 2011 and continues to cause high morbidity and about 5.0 to 50% mortality in ducklings. Currently there are no approved vaccines for the virus. This study aimed to assess the efficacy of a new vaccine created from the baculovirus and sigma C gene against NDRV. In this study, a recombinant baculovirus containing the sigma C gene was constructed, and the purified protein was used as a vaccine candidate in ducklings. The efficacy of sigma C vaccine was estimated according to humoral immune responses, cellular immune response and protection against NDRV challenge...
December 15, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27958330/highly-efficient-transduction-of-primary-adult-cns-and-pns-neurons
#10
Evgeny Levin, Heike Diekmann, Dietmar Fischer
Delivery and expression of recombinant genes, a key methodology for many applications in biological research, remains a challenge especially for mature neurons. Here, we report easy, highly efficient and well tolerated transduction of adult peripheral and central neuronal populations of diverse species in culture using VSV-G pseudo-typed, recombinant baculovirus (BacMam). Transduction rates of up to 80% were reliably achieved at high multiplicity of infection without apparent neuro-cytopathic effects. Neurons could be transduced either shortly after plating or after several days in culture...
December 13, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27955747/recombinant-expression-and-purification-of-functional-vascular-endothelial-growth-factor-121-in-the-baculovirus-expression-system
#11
Nastaran Mohseni, Ali Jahanian-Najafabadi, Fatemeh Kazemi-Lomedasht, Roghaye Arezomand, Mahdi Habibi-Anbouhi, Delavar Shahbazzadeh, Mahdi Behdani
OBJECTIVE: To express human vascular endothelial growth factor121 (VEGF121) in insect cells. METHODS: A gene construct containing VEGF was cloned in the pFastBac-HTA vector, followed by transformation in DH10BAC. The recombinant bacmid was then extracted, and transfected into Sf9 insect cells. The transfected cells were harvested, and then VEGF expression was confirmed by western blotting using specific antibodies. The tube formation assay was used for functional assessment of VEGF...
December 2016: Asian Pacific Journal of Tropical Medicine
https://www.readbyqxmd.com/read/27933303/purification-of-baculovirus-vectors-using-heparin-affinity-chromatography
#12
Md Nasimuzzaman, Danielle Lynn, Johannes Cm van der Loo, Punam Malik
Baculoviruses are commonly used for recombinant protein and vaccine production. Baculoviruses are nonpathogenic to vertebrates, have a large packaging capacity, display broad host and cell type tropism, infect both dividing and nondividing cells, and do not elicit strong immune or allergic responses in vivo. Hence, their use as gene delivery vehicles has become increasingly popular in recent years. Moreover, baculovirus vectors carrying mammalian regulatory elements can efficiently transduce and express transgenes in mammalian cells...
2016: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/27913752/expression-and-bioactivity-of-human-alpha-fetoprotein-in-a-bac-to-bac-system
#13
Bo Lin, Kun Liu, Wenting Wang, Wei Li, Xu Dong, Yi Chen, Yan Lu, Junli Guo, Mingyue Zhu, Mengsen Li
Alpha-fetoprotein (AFP) is an early serum growth factor in fetal embryonic development and hepatic oncogenesis. A growing number of investigations of AFP as a tumor-specific biomarker have concluded that AFP is an important target for cancer treatment. AFP also plays an immunomodulatory role in the treatment of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, myasthenia gravis, and thyroiditis. In an effort to support biochemical screening and drug design and discovery, we attempted to express and purify human AFP in a Bac-to-Bac system...
December 2, 2016: Bioscience Reports
https://www.readbyqxmd.com/read/27895582/introduction-of-an-n-glycosylation-site-into-udp-glucuronosyltransferase-2b3-alters-its-sensitivity-to-cytochrome-p450-3a1-dependent-modulation
#14
Tatsuro Nakamura, Naho Yamaguchi, Yuu Miyauchi, Tomoki Takeda, Yasushi Yamazoe, Kiyoshi Nagata, Peter I Mackenzie, Hideyuki Yamada, Yuji Ishii
Our previous studies have demonstrated functional protein-protein interactions between cytochrome P450 (CYP) 3A and UDP-glucuronosyltransferase (UGT). However, the role of carbohydrate chains of UGTs in the interaction with CYP is not well understood. To address this issue, we examined whether CYP3A1 modulates the function of UGT2B3 which lacks potential glycosylation sites. We also examined whether the introduction of N-glycosylation to UGT2B3 affects CYP3A-dependent modulation of UGT function. To introduce a potential glycosylation site into UGT2B3, Ser 316 of UGT2B3 was substituted with Asn by site-directed mutagenesis...
2016: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/27894868/the-91-205-amino-acid-region-of-acmnpv-orf34-ac34-which-comprises-a-potential-c3h-zinc-finger-is-required-for-its-nuclear-localization-and-optimal-virus-multiplication
#15
Jianxiang Qiu, Zhimin Tang, Meijin Yuan, Wenbi Wu, Kai Yang
During baculovirus infection, most viral proteins must be imported to the nucleus to support virus multiplication. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) orf34 (ac34) is an alphabaculovirus unique gene that is required for optimal virus production. Ac34 distributes in both the cytoplasm and the nuclei of virus-infected Sf9 cells, but contains no conventional nuclear localization signal (NLS). In this study, we investigated the nuclear targeting domains in Ac34. Transient expression assays showed that Ac34 localized in both the cytoplasm and the nuclei of Sf9 cells, indicating that no viral protein is required for Ac34 nuclear localization...
January 15, 2017: Virus Research
https://www.readbyqxmd.com/read/27888023/integrated-process-for-the-purification-and-immobilization-of-the-envelope-protein-domain-iii-of-dengue-virus-type-2-expressed-in-rachiplusia-nu-larvae-and-its-potential-application-in-a-diagnostic-assay
#16
María Emilia Smith, Alexandra Marisa Targovnik, Julieta Cerezo, María Alejandra Morales, María Victoria Miranda, Julián Rodríguez Talou
Dengue incidence has grown dramatically in the last years, with about 40% of the world population at risk of infection. Recently, a vaccine developed by Sanofi Pasteur has been registered, but only in a few countries. Moreover, specific antiviral drugs are not available. Thus, an efficient and accurate diagnosis is important for disease management. To develop a low-cost immunoassay for dengue diagnosis, in the present study we expressed the envelope protein domain III of dengue virus type 2 in Rachiplusia nu larvae by infection with a recombinant baculovirus...
November 23, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/27867046/efficient-production-of-an-avian-adeno-associated-virus-vector-using-insect-cell-baculovirus-expression-system
#17
Anping Wang, Yongjuan Wang, Shuang Wu, Weiyong Zuo, Changming Guo, Weiming Hong, Shanyuan Zhu
Recombinant avian adeno-associated virus (rAAAV) is a promising gene transfer vector for avian cells. Although rAAAV can be produced by co-transfection of HEK293 cells with three plasmids, both scalability and productivity of the transient transfection method can not meet the demand for large-scale in vivo experiments. In this study, a scalable rAAAV production method was established by using insect cell/baculovirus expression system. Three recombinant baculoviruses, namely BacARep, BacAVP and BacAGFP, were generated by transfection of Sf9 cells with the three plasmids expressing AAAV Rep genes, modified VP gene or the inverted terminal repeats-flanked green fluorescent protein (GFP) gene...
February 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/27860432/virus-like-particle-of-macrobrachium-rosenbergii-nodavirus-produced-in-spodoptera-frugiperda-sf9-cells-is-distinctive-from-that-produced-in-escherichia-coli
#18
Chare Li Kueh, Chean Yeah Yong, Seyedehsara Masoomi Dezfooli, Subha Bhassu, Soon Guan Tan, Wen Siang Tan
Macrobrachium rosenbergii nodavirus (MrNV) is a virus native to giant freshwater prawn. Recombinant MrNV capsid protein has been produced in Escherichia coli, which self-assembled into virus-like particles (VLPs). However, this recombinant protein is unstable, degrading and forming heterogenous VLPs. In the current study, MrNV capsid protein was produced in insect Spodoptera frugiperda (Sf9) cells through a baculovirus system. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the recombinant protein produced by the insect cells self-assembled into highly stable, homogenous VLPs each of approximately 40 nm in diameter...
November 14, 2016: Biotechnology Progress
https://www.readbyqxmd.com/read/27821590/heparan-sulfate-and-heparin-promote-faithful-prion-replication-in-vitro-by-binding-to-normal-and-abnormal-prion-proteins-in-protein-misfolding-cyclic-amplification
#19
Morikazu Imamura, Naoko Tabeta, Nobuko Kato, Yuichi Matsuura, Yoshifumi Iwamaru, Takashi Yokoyama, Yuichi Murayama
The precise mechanism underlying the conversion of normal prion protein (PrP(C)) into abnormal prion protein (PrP(Sc)) remains unclear. Protein misfolding cyclic amplification (PMCA), an in vitro technique used for amplifying PrP(Sc), results in PrP(Sc) replication that preserves the strain-specific characteristics of the input PrP(Sc); thus, PMCA mimics the process of in vivo PrP(Sc) replication. Previous work has demonstrated that in PMCA, nucleic acids are critical for PrP(Sc) amplification, but little information has been reported on glycosaminoglycan (GAG) participation in PrP(Sc) replication in vitro Here, we investigated whether GAGs play a role in the faithful replication of PrP(Sc) by using a modified PMCA performed with baculovirus-derived recombinant PrP (Bac-PrP) as a substrate...
December 16, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27818346/improved-cancer-specificity-in-psa-assay-using-aleuria-aurantia-lectin-coated-eu-nanoparticles-for-detection
#20
Henna Kekki, Mari Peltola, Sandra van Vliet, Chris Bangma, Yvette van Kooyk, Kim Pettersson
OBJECTIVES: The objective was to study the differences in PSA fucosylation obtained from LNCaP and PC-3 prostate cancer cell lines, seminal plasma PSA and recombinant precursor form of PSA expressed in baculovirus, using Aleuria aurantia lectin (AAL). The aim was to assess whether differences in fucosylation (Fucα1-6/3GlcNAc carbohydrates) of PSA either in urine, blood or tissue enable the discrimination of patients with prostate cancer (PCa) from benign prostatic hyperplasia (BPH) and young males...
January 2017: Clinical Biochemistry
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