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bombyx mori baculovirus

Susumu Katsuma, Ryuhei Kokusho
: The baculovirus VP39 is a major nucleocapsid protein essential for viral propagation. However, the critical domains or residues of the VP39 protein have not yet been identified. Here, we performed mutagenesis experiments of Bombyx mori nucleopolyhedrovirus (BmNPV) using 5-bromo-2' -deoxyuridine and isolated a BmNPV mutant that produced fewer occlusion bodies than the wild-type virus. This mutant also produced fewer infectious budded viruses (BVs), when compared with the wild-type virus in both cultured cells and B...
January 11, 2017: Journal of Virology
Shiho Tanaka, Haruka Endo, Satomi Adegawa, Shingo Kikuta, Ryoichi Sato
Bacillus thuringiensis produces Cry toxins, which are used as insecticides in sprays and in transgenic crops. However, little is known about the function of Cry toxin receptors and the mechanisms that determine their binding specificity and activity. In this study, the cRNAs of Bombyx mori ABC transporter C2 (BmABCC2), the toxin-binding region of cadherin-like receptor (BtR175-TBR), or aminopeptidase N1 (BmAPN1) were injected into Xenopus oocytes, and the Cry1Aa-dependent cation-selective pore formation activities of these receptors were analyzed using a two-electrode voltage clamp...
December 2016: FEBS Journal
Qiang Wang, Xiaoli Ju, Liang Chen, Keping Chen
Caspase-1 is one of the effector caspases in mammals that plays a central role in apoptosis. However, the lepidopteran caspase-1, especially the Bombyx mori caspase-1 (Bm-caspase-1), has not been investigated in detail. In this study, Bm-caspase-1 was identified from an expressed sequence tag database in B. mori by BLAST search. The open reading frame of Bm-caspase-1 contained 879 nucleotides and encoded 293 amino acids with a predicted molecular mass of 33 kDa. Bm-caspase-1 contained two consensus amino acid motifs of caspase cleavage sites, DEGDA and TETDG...
October 4, 2016: Zeitschrift Für Naturforschung. C, A Journal of Biosciences
Qi Qi, Lunguang Yao, Zhisheng Liang, Donghua Yan, Zhuo Li, Yadong Huang, Jingchen Sun
Human type II collagen is a macromolecular protein found throughout the human body. The baculovirus expression vector system is one of the most ideal systems for the routine production and display of recombinant eukaryotic proteins in insect, larvae, and mammalian cells. We use this system to express a full-length gene, human type II collagen cDNA (4257 bp), in cultured Spodoptera frugiperda 9 cells (Sf9), Bombyx mori cells, and silkworm larvae. In this study, the expression of human type II collagen gene in both insect cells and silkworm larvae was purified by nickel column chromatography, leading to 300-kDa bands in SDS-PAGE and western blotting indicative of collagen α-chains organized in a triple-helical structure...
December 2016: Molecular Genetics and Genomics: MGG
Y Zhang, Y Shi, H Yu, J Li, Y Quan, T Shu, Z Nie, Y Zhang, W Yu
Baculoviridae is a family of invertebrate viruses with large double-stranded DNA genomes. Proteins encoded by some late expression factor (lef ) genes are involved in the regulation of viral gene expression. Lef-9 is one of four transcription-specific Lefs, which are components of the virus-encoded RNA polymerase, and can initiate and transcribe late and very late genes. As a multifunctional protein encoded by the Bombyx mori nucleopolyhedrovirus (BmNPV), Lef-9 may be involved in the regulation of viral propagation...
2016: Acta Virologica
Ryuhei Kokusho, Yoshikazu Koh, Masaru Fujimoto, Toru Shimada, Susumu Katsuma
Bombyx mori nucleopolyhedrovirus (BmNPV) orf5 (Bm5) is a core gene of lepidopteran baculoviruses and encodes the protein with the conserved amino acid residues (DUF3627) in its C-terminus. Here, we found that Bm5 disruption resulted in lower titers of budded viruses and fewer numbers of occlusion bodies (OBs) in B. mori cultured cells and larvae, although viral genome replication was not affected. Bm5 disruption also caused aberrant expression of various viral genes at the very late stage of infection. Immunocytochemical analysis revealed that BM5 localized to the nuclear membrane...
November 2016: Virology
Shumin Liu, Panli Zhang, Hong-Sheng Song, Hai-Sheng Qi, Zhao-Jun Wei, Guozheng Zhang, Shuai Zhan, Zhihong Liu, Sheng Li
The Hippo pathway, which was identified from genetic screens in the fruit fly, Drosophila melanogaster, has a major size-control function in animals. All key components of the Hippo pathway, including the transcriptional coactivator Yorkie that is the most critical substrate and downstream effector of the Hippo kinase cassette, are found in the silkworm, Bombyx mori. As revealed by microarray and quantitative real-time PCR, expression of Hippo pathway genes is particularly enriched in several mitotic tissues, including the ovary, testis, and wing disc...
2016: International Journal of Biological Sciences
Rui Guo, Guangli Cao, Yuexiong Zhu, Dhiraj Kumar, Renyu Xue, Yahong Lu, Xiaolong Hu, Chengliang Gong
Bombyx mori bidensovirus (BmBDV) was previously termed as Bombyx mori densovirus type 2 and later it was reclassified in the new genus bidensovirus of the new family Bidnaviridae. The genome of BmBDV Zhenjiang isolate (BmBDV-Z) consists of two non-homologous single-stranded linear DNA molecules VD1 and VD2 which are encapsidated into separate virion. To investigate the infectivity of BmBDV DNA, recombinant plasmids pGEM-VD1 inserted with VD1 genome were transfected into the BmN cells of silkworm. Structural proteins of BmBDV were detected with Western blot and immunofluorescence assay, which indicates pGEM-VD1 replicated in the transfected BmN cells and viral proteins were also expressed...
October 2016: Current Microbiology
Cui Zhao, Chen Zhang, Bin Chen, Yanghui Shi, Yanping Quan, Zuoming Nie, Yaozhou Zhang, Wei Yu
A DNA-binding protein (DBP) [GenBank accession number: M63416] of Bombyx mori nuclear polyhedrosis virus (BmNPV) has been reported to be a regulatory factor in BmNPV, but its detailed functions remain unknown. In order to study the regulatory mechanism of DBP on viral proliferation, genome replication, and gene transcription, a BmNPV dbp gene knockout virus dbp-ko-Bacmid was generated by the means of Red recombination system. In addition, dbp-repaired virus dbp-re-Bacmid was constructed by the means of the Bac to Bac system...
2016: PloS One
Ming-Ya Cao, Xiu-Xiu Kuang, Hai-Qing Li, Xue-Jiao Lei, Wen-Fu Xiao, Zhan-Qi Dong, Jun Zhang, Nan Hu, Ting-Ting Chen, Cheng Lu, Min-Hui Pan
Pathogen-inducible promoters have been studied extensively and widely used in resistance breeding and gene therapy. However, few reports have been published that explore the efficacy of Bombyx mori nucleopolyhedrovirus (BmNPV)-inducible promoters in antiviral research in the Bombyx mori (Lepidoptera). Here, we screened BmNPV promoters (VP1054, P33, Bm21, Bm122, 39K, P143, and P6.9) and found that the 39K promoter had the highest BmNPV-induced transcriptional activity by dual-luciferase reporter assays system...
August 10, 2016: Journal of Biotechnology
Toshihiro Nagamine, Yasushi Sako
Although phylogenic analysis often suggests co-evolutionary relationships between viruses and host organisms, few examples have been reported at the microevolutionary level. Here, we show a possible example in which a species-specific anti-viral response may drive phylogenic divergence in insect virus evolution. Two baculoviruses, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV), have a high degree of DNA sequence similarity, but exhibit non-overlapping host specificity...
2016: PloS One
Feifei Yin, Ruikun Du, Wenhua Kuang, Guang Yang, Hualin Wang, Fei Deng, Zhihong Hu, Manli Wang
Fibroblast growth factor (FGF) is found throughout multicellular organisms; however, fgf homologs (vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of vFGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV), involves accelerated killing of infected larvae by both viruses. The vFGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites...
June 2016: Virologica Sinica
Kazuhiro Iiyama, Jae Man Lee, Tuneyuki Tatsuke, Hiroaki Mon, Takahiro Kusakabe
Baculovirus-Bombyx mori protein expression system has mainly been used for translation of eukaryotic proteins. In contrast, information pertaining to bacterial protein expression using this system is not sufficient. Therefore, recombinant nucleases from Serratia liquefaciens (rSlNucAs) were expressed in a Baculovirus-B. mori protein expression system. rSlNucAs containing the native signal peptide (rSlNucA-NSP) or silkworm 30-K signal peptide (rSlNucA-30K) at the NH2-terminus were constructed to enable secretion into the extracellular fraction...
June 2016: Molecular Biotechnology
Xingjian Liu, Yonglong Wei, Yinü Li, Haoyang Li, Xin Yang, Yongzhu Yi, Zhifang Zhang
The silkworm baculovirus expression system is widely used to produce recombinant proteins. Several strategies for constructing recombinant viruses that contain foreign genes have been reported. Here, we developed a novel defective-rescue BmNPV Bacmid (reBmBac) expression system. A CopyControl origin of replication was introduced into the viral genome to facilitate its genetic manipulation in Escherichia coli and to ensure the preparation of large amounts of high quality reBmBac DNA as well as high quality recombinant baculoviruses...
2016: PloS One
Qiang Wang, Yang Zhou, Keping Chen, Xiaoli Ju
Baculovirus expression vector systems (BEVSs) have been widely used for recombinant protein production. Many studies have tried to improve the recombinant protein production by either modification of BEVSs vectors or by cell line selection. In this study, using a modified shRNA vector, we established a stable Bombyx mori cell line that significantly inhibits expression of caspase-1 after selection. We further compared cell proliferation and viability between caspase-1-suppressed cells and control cells, and found that there is no significant difference in these stable cell lines...
May 2016: Molecular Biotechnology
Zhan-Qi Dong, Ting-Ting Chen, Jun Zhang, Nan Hu, Ming-Ya Cao, Fei-Fan Dong, Ya-Ming Jiang, Peng Chen, Cheng Lu, Min-Hui Pan
Although current antiviral strategies can inhibit baculovirus infection and decrease viral DNA replication to a certain extent, novel tools are required for specific and accurate elimination of baculovirus genomes from infected insects. Using the newly developed clustered regularly interspaced short palindromic repeats/associated protein 9 nuclease (CRISPR/Cas9) technology, we disrupted a viral genome in infected insect cells in vitro as a defense against viral infection. We optimized the CRISPR/Cas9 system to edit foreign and viral genome in insect cells...
June 2016: Antiviral Research
Alexandra M Targovnik, Mariana B Arregui, Lautaro F Bracco, Nicolas Urtasun, Maria F Baieli, Maria M Segura, Maria A Simonella, Mariela Fogar, Federico J Wolman, Osvaldo Cascone, Maria V Miranda
In Biotechnology, the expression of recombinant proteins is a constantly growing field and different hosts are used for this purpose. Some valuable proteins cannot be produced using traditional systems. Insects from the order Lepidoptera infected with recombinant baculovirus have appeared as a good choice to express high levels of proteins, especially those with post-translational modifications. Lepidopteran insects, which are extensively distributed in the world, can be used as small protein factories, the new biofactories...
2016: Current Pharmaceutical Biotechnology
Ladislav Roller, Daniel Čižmár, Zuzana Gáliková, Branislav Bednár, Ivana Daubnerová, Dušan Žitňan
Trissin has recently been identified as a conserved insect neuropeptide, but its cellular expression and function is unknown. We detected the presence of this neuropeptide in the silkworm Bombyx mori using in silico search and molecular cloning. In situ hybridisation was used to examine trissin expression in the entire central nervous system (CNS) and gut of larvae, pupae and adults. Surprisingly, its expression is restricted to only two pairs of small protocerebral interneurons and four to five large neurons in the frontal ganglion (FG)...
June 2016: Cell and Tissue Research
Konstantinos Ioannidis, Luc Swevers, Kostas Iatrou
In this study, we have deleted the lef8 gene of the baculovirus BmNPV, which encodes one of the viral RNA polymerase subunits, in order to create a knockout bacmid, Δlef8, directing cytopathology-free single-cell infections for gene transduction and recombinant protein production. However, while removal of the complete lef8 ORF produced the expected phenotype, it also affected the function of the closely linked essential gene orf40, thus hampering the mutant bacmid rescue in cultured Bombyx cells expressing recombinant LEF8...
March 2016: Journal of General Virology
Katsuhisa Innami, Takahiro Aizawa, Toshihiro Tsukui, Susumu Katsuma, Shigeo Imanishi, Hideki Kawasaki, Masashi Iwanaga
Bombyx mori-derived cell lines are generally used for Bombyx mori nucleopolyhedrovirus (BmNPV)-based baculovirus expression vector system (BEVS). However, almost all of the B. mori-derived cell lines are persistently infected with Bombyx mori macula-like virus (BmMLV). In this study, nontarget mammalian cell lines were exposed to BmMLV, and their susceptibility was investigated. Real-time PCR showed that viral RNA in virus-inoculated nine mammalian cell lines decreased sharply at 7 days postinfection. Also, there was no significant effect on cell viability of mammalian cells after inoculation with BmMLV...
March 2016: Journal of Virological Methods
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