• Title/Summary/Keyword: AcNPV

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Effect of 1-deoxynojirimycin on the Replication of Baculoviruses, Bombyx Mori Nucleopolyhedrovirus and Autographa Californica Multiple Nucleopolyhedrovirus

  • Kang, Kyung-Don;Park, Joo-Sung;Cho, Yong-Seok;Park, Young-Shik;Lee, Jae-Yeon;Hwang, Kyo-Yeol;Yuk, Won-Jeong;Kamita, Shizuo George;Suzuki, Koichi;Seong, Su-Il
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.123-128
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    • 2011
  • 1-Deoxynojirimycin (DNJ) is an alkaloid that is found at relatively high concentrations in mulberry leaf and tissues of the silkworm, $Bombyx$ $mori$. DNJ is a well known inhibitor of ${\alpha}$-glucosidase, an enzyme that is involved in the early stages of the $N$-linked glycoprotein synthesis pathway. ${\alpha}$-Glucosidase activity in the cell extract from $B.$ $mori$-derived Bm5 cells showed approximately 40-fold less sensitivity to DNJ than ${\alpha}$-glucosidase activity in the cell extract from $Spodoptera$ $frugiperda$-derived Sf9 cells. The replication of $B.$ $mori$ nucleopolyhedrovirus (BmNPV) was not inhibited when it was propagated in BmN cells that were grown in medium containing up to 10 mM DNJ. In contrast, the replication of $Autographa$ $californica$ multiple NPV (AcMNPV) was reduced by 67% when it was propagated in Sf9 cells that were grown in medium containing 10 mM DNJ. The viability of Bm5 and Sf9 cells that were grown in medium containing up to 10 mM DNJ was not affected. Our results suggested that the reduced replication of AcMNPV was the result of the higher sensitivity of ${\alpha}$-glucosidase activity in Sf9 cells to DNJ.

Construction of a Transposon-mediated Baculovirus Vector Hanpvid and a New Cell Line for Expressing Barnase

  • Qin, Qin;Liu, Ying-Le;Zhu, Ying;Li, Shun-Yi;Qi, Yi-Peng
    • BMB Reports
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    • v.38 no.1
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    • pp.41-48
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    • 2005
  • In this study we developed the transposon-mediated shuttle vector 'Hanpvid', which composed of HaNPV (Heliothis armigera nuclear polyhedrosis virus) genomic DNA and a transposon cassette from Bacmid of Bac-to-Bac system. Hanpvid replicates in E. coli in the same way as Bacmid and retains infective function in cotton bollworm cells (Hz-AM1). Using Hanpvid we constructed a recombinant virus, which could infect Hz-AM1 cells and generate recombinant HaNPV (rHa-Bar) containing the barnase gene, a ribonuclease gene from Bacillus amyloliquefaciens. Since the expression vector carrying barnase gene cannot replicate in the absence of barstar, a specific inhibitor of barnase, we constructed a new cotton bollworm cell line (AM1-NB) using the marker rescue method. In AM1-NB barstar was integrated into the cellular chromosome to sustain the replication of rHa-Bar. To screen out recombinant HaNPV for potential use as biopesticide, Hz-AM1 and AM1-NB cell lines were infected with rHa-Bar, respectively. The results obtained indicate that Viral progenies in AM1-NB were 23 and 160 times greater than those in Hz-AM1 48 h and 72 h after infection, respectively. With additional insertion of the polyhedron gene from AcNPV (Autographa californica nuclear polyhedrosis virus) into the Hanpvid genome, rHa-Bar regained the polyhedron phenotype and its pest-killing rate greatly improved. Toxic analysis showed that the lethal dosages ($LD_{50}$) and the lethal time(s) ($LT_{50}$) of rHa-Bar were reduced by 20% and 30%, respectively, compared to wt-HaNPV in the third instar larvae of cotton bollworm. This study shows that in AM1-NB barnase can be effectively produced and used as pest-killing agent for the biological control of cotton pests.

Control and Analysis of Vienna Rectifier Used as the Generator-Side Converter of PMSG-based Wind Power Generation Systems

  • Zhao, Hongyan;Zheng, Trillion Q.;Li, Yan;Du, Jifei;Shi, Pu
    • Journal of Power Electronics
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    • v.17 no.1
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    • pp.212-221
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    • 2017
  • Permanent-Magnet Synchronous Generators (PMSGs) are used widely in Wind Power Generation Systems (WPGSs), and the Vienna rectifier was recently proposed to be used as the generator-side converter to rectify the AC output voltage in PMSG-based WPGS. Compared to conventional six-switch two-level PWM (2L-PWM) converters, the Vienna rectifier has several advantages, such as higher efficiency, improved total harmonic distortion, etc. The motivation behind this paper is to verify the performance of direct-driven PMSG wind turbine system based-Vienna rectifier by using a simulated direct-driven PMSG WPGS. In addition, for the purpose of reducing the reactive power loss of PMSGs, this paper proposes an induced voltage sensing scheme which can make the stator current maintain accurate synchronization with the induced voltage. Meanwhile, considering the Neutral-Point Voltage (NPV) variation in the DC-side of the Vienna rectifier, a NPV balancing control strategy is added to the control system. In addition, both the effectiveness of the proposed method and the performance of the direct-driven PMSG based-Vienna rectifier are verified by simulation and experimental results.

Partial genomic sequence of baulovirus associated with white spot syndrome (WSBV) isolated from penaeid shrimp P. chinensis (대하새우로부터 분리한 WSBV의 게놈서열 분석)

  • Kim, Chong-Kyung;Sohn, Sang-Gyu;Heo, Moon-Soo;Lee, Tae-Ho;Jun, Hong-Ki;Jang, Kyung-Lib
    • Journal of fish pathology
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    • v.10 no.2
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    • pp.87-95
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    • 1997
  • Baculovirus associated with white spot syndrome (WSBV) is the causative agent of a disease with high mortalities and causes severe damage to shrimp cultures. In this study, we analyzed a recombinant clone (E3) obtained from a viral genomic library to characterize the causative agent in diseased shrimp Penaeus chinensis with white spot syndrome. According to the analysis of nucleotide sequence of E3, this clone did not showed considerable sequence homology with those of other known viruses, including baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV), indicating that WSBV is a novel virus causing a serious disease in P. chinensis. Based on the sequence of E3 clone, a pair of PCR primers was designed. After 30 cycles of amplification, a specific product of the expected size was detected only if the total nucleic acids extracted from the diseased shrimp was used as a template DNA, suggesting that this method can be used to diagnose the virus infection in diseased shrimp.

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Construction of a Transgenic Silkworm Carrying the Fibroin Gene of the Japanese Oak Silkworm, Antheraea yamamai

  • Park, Kwang-Ho;Kang, Seok-Woo;Hwang, Jae-Sam;Goo, Tea-Won;Yun, Eun-Young;Lee, Sang-Mong;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.1
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    • pp.49-55
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    • 2003
  • We describe the generation of transgenic silkworm that carrying the chimeric fibroin light chain (L-chain) gene. Previously, we have cloned the complete fibroin L-chain gene from the silkworm Baekok-Jam, Bombyx mori, and the complete fibroin gene from the oak silkworm, Antheraea yamamai. The 444 bp repetitive sequence of A. yamamai fibroin gene was inserted into the exon 6 of B. mori fibroin L-chain gene to produce chimeric fibroin L-chain gene. The chimeric fibroin L-chain gene was cloned into the polyhedrin gene site of Autographa californica nuclear polyhedrosis virus (AcNPV) to yield a recombinant baculovirus as a fibroin gene targeting vector, One-day-old fifth instar female silkworm larvae were injected with the recombinant baculovirus and then mated with normal male moths. Genomic DNA from their progenies was extracted and screened for the desired targeting event by using PCR and Southern blot analysis. The analysis showed that the chimeric fibroin gene had intergrated into the L-chain gene on the genome by homologous recombination and was transmitted through generations. The transgenic silkworm carrying the chimeric fibroin gene were approximately 43.2% in $F_2$ generation, and the silkworms synthesized the fusion protein in cocoons layer.

Anticancer and Antiviral Activity of Chlorine Dioxide by Its Induction of the Reactive Oxygen Species (이산화염소의 활성산소 생성 유도에 의한 항암 및 항바이러스 활성)

  • Kim, Yonggyun;Kumar, Sunil;Cheon, Wonsu;Eo, Hyunji;Kwon, Hyeok;Jeon, Yongho;Jung, Jinboo;Kim, Wook
    • Journal of Applied Biological Chemistry
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    • v.59 no.1
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    • pp.31-36
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    • 2016
  • Chlorine dioxide has been used for a disinfectant by exhibiting antimicrobial activity and is also potent to kill insect pests infesting stored grains. This study aimed to extend the usefulness of chlorine dioxide with respect to anticancer and antiviral activities. Cytotoxicity of chlorine dioxide was assessed against five different human cancer cell lines. Chlorine dioxide exhibited significant cytotoxicity against two breast cancer cell lines (MCF-7, MDA-MB-231) and three colorectal cancer cell lines (LoVo, HCT-116, SW-480). This cytotoxicity appeared to be associated with the capacity of chlorine dioxide to induce the production of reactive oxygen species (ROS). Compared to control insect cell lines, the cancer cell lines possessed much higher levels of ROS. On the other hand, a treatment of an antioxidant, vitamin E, significantly reduced the cytotoxicity, suggesting that the cytotoxicity was induced by high levels of ROS production. Chlorine dioxide exhibited antiviral activity against different viruses. A baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV), is a dsDNA insect virus and lost its viral activity to form polyhedral viral particles in response to chlorine dioxide. The antiviral activity against AcNPV was dependent on the incubation time with chlorine dioxide. Tobacco mosaic virus is a ssRNA plant virus and was reduced in its population after exposure to chlorine dioxide along with significant decrease of viral symptoms. These results indicate that chlorine dioxide possesses anticancer and antiviral activities probably due to its inducing activity of ROS production.

Expression of Human Thrombopoietin in Insect Cells by Polyhedrin-gp64 Dual Promoter-Based Baculovirus Vector System

  • Koh, Yeo-Wook;Park, Sang-Kyu;Kim, Kun-Soo;Kim, Seong-Ryong;Lee, Jeong-Kug;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.318-322
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    • 1999
  • A new baculovirus transfer vector (pPGP404) was constructed to increase the expression level of human thrombopoietin (hTPO) in insect cells. In pPGP404, hTPO was cloned next to the AcNPV polyhedrin-gp64 dual promoter and the leader sequence of hTPO was substituted with that of gp64. A recombinant baculovirus, AcPGP404, was constructed by using pPGP404 as a transfer vector. hTPO was expressed in AcPGP404-infected TN5 cells and it was observed that the expression levels of hTPO in TN5 cells increased three-fold ($6.0 {\mu}g/ml^{-1}$) compared to the level expressed under the control of the polyhedrin single promoter. These results indicate that the polyhedrin-gp64 dual promoter system would be useful for expression in large quantities of recombinant proteins in insect cells.

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A Control Scheme for Quality Improvement of Input-Output Current of Small DC-Link Capacitor Based Three-Level NPC Inverters (소용량 직류단 커패시터를 가지는 3-레벨 NPC 인버터의 입-출력 전류 품질 향상을 위한 제어 기법)

  • In, Hyo-Chul;Kim, Seok-Min;Park, Seong-Soo;Lee, Kyo-Beum
    • The Transactions of the Korean Institute of Power Electronics
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    • v.22 no.4
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    • pp.369-372
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    • 2017
  • This paper presents a control scheme for three-level NPC inverters using small DC-link capacitors. To reduce the inverter system volume, the film capacitor with small capacitance is a promising candidate for the DC-link. When small capacitors are applied in a three level inverter, however, the AC ripple component increases in the DC-link NPV (neutral point voltage). In addition, the three-phase input grid currents are distorted when the DC-link capacitors are fed by diode rectifier. In this paper, the additional circuit is applied to compensate for small capacitor systems defect, and the offset voltage injection method is presented for the stabilization in NPV. These two proposed processes evidently ensure the quality improvement of the input grid currents and output load currents. The feasibility of the proposed method is verified by experimental results.

Effects of Insect Hormones on the Replication of Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Yi, Yong-Zhu;Xiao, Qing-Li;He, Jia-Lu;Zhou, Ya-Jing;Zhang, Yuan-Xing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.2
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    • pp.137-141
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    • 2002
  • An experimental study was undertaken to quantify the effects of insect hormones on the replication of nucleopolyhedrovirus (NPV). The results demonstrated that TCID/ sub 50/ at 72 h post-infection (hpi) rose systematically from 0.55$\times$10$^{8}$ /m1, for untreated cells, up to 1.67$\times$10$^{8}$ / ml at 3$\mu$g/ml, then dropped down to 1.45$\times$10$^{8}$ /m1 at 4 $\mu$g/ml, by adding ecdysone to the culture medium for Bm-N cells infected with a wild-type Bambyx mori. nucleopolyhedrovirus (BmNPV). The optimum enhancement of about 3 times on budded virus (BV) titer at 72 hpi was given at 3 $\mu$g/ml of ecdysone. While the polyhedra number had no obvious variation within the range of concentrations from 0 to 4 $\mu$g/ml. By addition of juvenile hormone analogue (JHA) into the media with this concentration range, the BmNPV TCID/ sub 50/ and polyhedra number at 72 hpi did not show significant changes. Also, the addition of either 3 $\mu$g/ml of ecdysone or 3 $\mu$g/ml of JHA to the culture media did not appear to affect the TCID/ sub 50/ and polyhedra number significantly in infected Sf-21 cells with the autographa californica nucleopolyhedrovirus (AcMNPV).