• BMB Reports
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• 제34권4호
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• pp.371-378
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• 2001

The Glycoprotein D (gD) gene of the HSV-1 strain F was cloned, sequenced, recombinated into the HcNPV (Hyphantria cunea nuclear polyhedrosis virus) expression vector and expressed in insect cells. The gD gene was located in the 6.43 kb BamHI fragment of the strainF. The open reading frame (ORF) of the gD gene was 1,185 by and codes 394 amino acid residues. Recombinant baculoviruses, GD-HcNPVs, expressing the gD protein were constructed. Spodoptera frugiperda cells, infected with the recombinant virus, synthesized a matured gX-gD fusion protein with an approximate molecular weight of 54 kDa and secreted the gD proteins into the culture media by an immunoprecipitation assay The fusion gD protein was localized on the membrane of the insect cells, seen by using an immunofluorescence assay The deduced amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. These results indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins.

• BMB Reports
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• 제37권6호
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• pp.735-740
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• 2004

Hepatitis C virus (HCV) is a causal agent of the chronic liver infection. To understand HCV morphogenesis, we studied the assembly of HCV structural proteins in insect cells. We constructed recombinant baculovirus expression vectors consisting of either HCV core alone, core-E1, or core-E1-E2. These structural proteins were expressed in insect cells and were examined to assemble into particles. Neither core-E1 nor core-E1-E2 was capable of assembling into virus-like particles (VLPs). It was surprising that the core protein alone was assembled into core-like particles. These particles were released into the culture medium as early as 2 days after infection. In our system, HCV structural proteins including envelope proteins did not assemble into VLPs. Instead, the core protein itself has the intrinsic capacity to assemble into amorphous core-like particles. Furthermore, released core particles were associated with HCV RNA, indicating that core proteins were assembled into nucleocapsids. These results suggest that HCV may utilize a unique core release mechanism to evade the hosts defense mechanism, thus contributing to the persistence of HCV infection.

• 농약과학회지
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• 제5권4호
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• pp.11-19
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• 2001

곤충의 난황형성에는 각종호르몬들이 먹이 등과 함께 매우 중요하게 작용하고 있으며, 이러한 호르몬들 중에서는 신경분비호르몬, 유약호르몬 및 엑디스테로이드 등이 주작용을 한다. 본 총설에서는 곤충호르몬을 포함한 곤충성장조절제가 곤충의 난황형성에 작용하여 어떤 영향을 주는지에 대하여 살펴보고 종합적 해충관리 인자로서 이용 가능성을 조사하였다. 곤충의 난황형성은 일반적으로 뇌에서 분비하는 신경분비호르몬의 작용과 난소에서 분비되는 엑디스테로이드 및 알라타체에서 분비되는 유약호르몬이 지방체를 자극하여 난황소전구물질을 합성하게 하며, 혈림프로 분비된 난황소 전구물질은 난모세포로 흡수되어 난발육이 이루어진다. 곤충호르몬을 포함한 곤충성장조절제는 이러한 난황형성과정 중에 각종 호르몬분비 기관(알라타체와 난소 등) 및 여러 가지 생리기관(지방체, 중장상피 그리고 침샘 등)에 작용하여 용해, 파괴 및 세포독성을 나타내 정상적인 생식작용을 하지 못하도록 한다. 따라서 곤충의 종류에 따라서는 포유동물에 독성이 낮은 곤충호르몬을 포함한 곤충성장조절제를 종합적 해충관리의 일환으로 사용함으로서 효과적인 해충방제가 가능할 것으로 보인다.

• Journal of Microbiology
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• 제45권2호
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• pp.133-138
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• 2007

This study addresses the susceptibility of Spodoptera frugiperda (Sf9 and Sf21), Trichoplusia ni (Hi5), and S. exigua (Se301) cells to the Bombyx mori nucleopolyhedrovirus (BmNPV). Although these cells have classically been considered nonpermissive to BmNPV, the cytopathic effect, an increase in viral yield, and viral DNA synthesis by BmNPV were observed in Sf9, Sf21, and Hi5 cells, but not in Se301 cells. Very late gene expression by BmNPV in these cell lines was also detected via ${\beta}-galactosidase$ expression under the control of the polyhedrin promoter. Sf9 cells were most susceptible to BmNPV in all respects, followed by Sf21 and Hi5 cells in decreasing order, while the Se301 cells evidenced no distinct viral replication. This particular difference in viral susceptibility in each of the cell lines can be utilized for our understanding of the mechanisms underlying the host specificity of NPVs.

• Molecules and Cells
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• 제28권3호
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• pp.155-165
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• 2009

The newly sequenced complete mitochondrial genome of the brown marmorated stink bug, Halyomorpha halys($St{\aa}l$) (Hemiptera: Pentatomidae), is a circular molecule of 16,518 bp with a total A+T content of 76.4% and two extensive repeat regions in A+T rich region. Nucleotide composition and codon usage of H. halys are about average when compared with values observed in 19 other published hemipteran mitochondrial genomes. Phylogenetic analyses using these 20 hemipteran mitochondrial genomes support the currently accepted hypothesis that suborders Heteroptera and Auchenorrhyncha form a monophyletic group. The mitochondrial gene arrangements of the 20 genomes are also consistent with our results.

• Animal cells and systems
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• 제15권2호
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• pp.169-177
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• 2011

In August 2002, a heavy rainfall (445 mm in total for 5 consecutive days) resulted in a catastrophic flood, and it completely washed away the benthic fauna from the mainstream channel of the Gapyeong stream, a typical mid-sized stream in the central Korean peninsula. This study was to investigate the recovery patterns of aquatic insect communities that were damaged by the flood. Aquatic insects were sampled quantitatively using a Surber sampler ($50{\times}50$ cm, 1 riffle and 1 pool/run habitats per site) from three sites (4th-6th order) of the Gapyeong stream prior to 2000 and seasonally after the flood event from 2003 to 2006. Before the flood in the reference year (2000), a total of 77 species of aquatic insects were collected, whereas after the flood 47 species (2003), 51 species (2004), 64 species (2005) and 55 species (2006) were collected from the whole sampling sites. The aquatic insect density decreased to 26.85% (2003), 90.25% (2004), 52.53% (2005) and 54.95% (2006) of that recorded in the reference year. Although approximately 70% of the aquatic insect fauna has recovered since the flood event, the species composition in the most recent year differed substantially (similarity ca. 50%). On the other hand, the compositions of functional groups have not significantly changed. Aquatic insect communities at the riffle sites were affected more profoundly than those at the pool/run sites. The aquatic insect communities at the upstream site recovered more rapidly than those at the downstream sites.

• BMB Reports
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• 제31권3호
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• pp.281-286
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• 1998

Hepatitis C virus (HCV) core proteins from two different isolates (HCV-1 and HCV-RH) were expressed in Spotioptera Jrugiperda (Sf9) insect cells. The RH core consisted of two major species of proteins (21 kDa and 19 kDa). On the other hand, the HCV-1 core was approximately 16 kDa in a SDS-PAGE gel. Both core proteins were phosphorylated in vivo on serine residues. Furthermore, the RH core but not HCV-1 core formed dimers, indicating that the protein conformation of the core in these two isolates is dfferent from one another. Immunofluorescence studies showed that the RH core was present in the cytoplasm, whereas the HCV-1 core was localized predominantly to the nucleus in recombinant baculovirus-infected insect cells. Since the major difference between the two isolates is the codon 9 of the core protein, a single amino acid substitution appears to play a major role in the protein conformation and these properties may reflect the different biological functions of core proteins in HCV-infected cells.

• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.200-203
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• 1994

Spodoptera frugiperda IPLB-SF-21-AE cells were cultivated in a spin-filter bioreactor with continuous perfusion for the recombinant $\beta$-galactosidase production. At the perfusion rate of 0.06 $hr^{-1}$, the maximum cell density of insect cells in this bioreactor system reached 3.5$\times$$l0^6$ viable cells/ml using the Grace media containing 5% FBS and 0.3% Pluronic F-68. The recombinant $\beta$-galactosidase production of 8, 100 units per reactor volume was also achieved at this perfusion rate.

• BMB Reports
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• 제39권3호
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• pp.263-269
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• 2006

In the genome of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, open reading frame 39 (Ha39) is the only gene predicted to encode an RNA recognition protein. Computer analysis revealed that Ha39 homologues were found in 15 NPVs, but not in GVs. Its transcripts were detected from 3 through 72 hours post infection (h p.i.) using RT-PCR and Northern blot analysis. The protein was detected in infected-cell lysates from 6 h p.i. Western blot assay of ODV and BV preparations revealed that Ha39 encodes a structural protein associated with BVs. Additionally, immunofluorescence microscopy demonstrated that the protein was present within cytoplasm in virus-infected cells, but not in the nuclear region.

• International Journal of Industrial Entomology and Biomaterials
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• 제36권1호
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• pp.15-24
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• 2018

N-glycosylation is an important posttranslational modification that results in a variety of biological activities, structural stability, and protein-protein interactions. There are still many mysteries in the structure and function of N-glycans, and detailed elucidation is necessary. Baculovirus expression system (BES) is widely used to produce recombinant glycoproteins, but it is not suitable for clinical use due to differences in N-glycan structure between insects and mammals. It is necessary to develop adequate model glycoproteins for analysis to efficiently alter the insect-type N-glycosylation pathway to human type. The previous research shows the recombinant alpha 1-acid glycoprotein (${\alpha}1AGP$) secreted from silkworm cultured cells or larvae is highly glycosylated and expected to be an excellent research candidate for the glycoprotein analysis expressed by BES. Therefore, we improved the ${\alpha}1AGP$ to be a better model for studying glycosylation. The modified ${\alpha}1AGP$ (${\alpha}1AGP{\Delta}$) recombinant protein was successfully expressed and purified by using BES, however, the expression level in silkworm cultured cells and larvae were lower than that of the ${\alpha}1AGP$. Subsequently, we confirmed the detailed profile of N-glycan on the ${\alpha}1AGP{\Delta}$ by LS/MS analysis the N-glycan structure at each glycosylation site. These results indicated that the recombinant ${\alpha}1AGP{\Delta}$ could be usable as a better model glycoprotein of N-glycosylation research in BES.