• Journal of Sericultural and Entomological Science
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• v.32 no.1
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• pp.49-57
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• 1990

To investigate the pathway of Nuclear Polyhedrosis Virus(NPV) of Bombyx mori in early stage infection of 2nd instar larva, the larval tissues were observed under electron microscope at interval of 6, 12, 24, and 48 hours after virus inoculation. The results are as follows. 1. The intact and enveloped nucleocapsids released from the polyhedra protein in the gut lumen apparently entered with the microvilli. 2. Virus progenies were observed in columnar cell nuclei 24 hours after inoculation, but polyhedra was not seen. The enveloped virus was observed in some of the intercellular spaces between mid-gut cells. 3. Many enveloped virus particles appeared in the basement membrane. These enveloped virus particles passed the basement membrane and gathered at blood cells in heomocoeal. 4. The NPV muliplicates in nuclei of the blood cells and the tracheal cells normally.

• Korean Journal of Animal Reproduction
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• v.19 no.1
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• pp.35-41
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• 1995

In this study, we have tested whether the retrovirus vector system is applicable in transgenic cattle production. To overcome low infectivity of currently available retrovirus vector system we have directly microinjected retrovirus-producing cells into the perivitelline space of the day 1.5 embryos. The virus-producing cell line was designed to release replication-defective retrovirus encapsidated with Gibbon ape leukemia virus (GaLV) envelope protein. E. coli LacZ gene was used as a marker gene to facilitate evaluation of the transgene expression and X-gal staining at morula or blastocyst stage resulted in expression of E. coli LacZ gene The results in these experiments were summarized as follows : 1. The lowest concentration of polybrene necessary for efficient virus infection was Sf' g/ml. 2. Development rate from day 1.5 embryos microinjected with virus-producing cells to the morulae /blastocysts was 29%. 3. 21% of the morulae /blastocysts were LacZ+. 4. There was no evidence that the retrovirus-producing cells used in this study produced replication-competent retrovirus.

• Korean journal of applied entomology
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• v.34 no.3
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• pp.218-223
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• 1995

A nuclear polyhedrosis virus isolated from the oriental tobacco budworm larvae, Helicoverpa assulta (Guenee) was characterized by electron microscopy, SDS-PAGE, restriction endonuclease analysis and cross infectivity. The shape of a polyhedron was $1.0\mu\textrm{m}$ in average with icosahdral outline, and the virus particle was $65nm\times300nm$ in average with rod-shape. The nuclear polyhedrosis virus was contained a single nucleocapsid within a viral envelope embedded in a polyhedron. The polyhedral protein was composed of a single polypeptide with a M.W. of 31 Kd. The genome size of the virus by restriction endonuclease analysis was about 120 Kb. Among several nuclear polyhedrosis viruses, the nuclear polyhedrosis virus from Helicoverpa assulta (HaNPV) and Autographa california nuclear polyhedrosis virus (AcNPV) were infected the oriental tobacco budworm larvae.

• Journal of Microbiology
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• v.34 no.1
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• pp.7-14
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• 1996

Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

• Journal of Microbiology
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• v.35 no.1
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• pp.72-77
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• 1997

The baculovirus gp64 glycoprotein is a major component of the envelope protein of budded virus (BV). It has been shown that the gp64 glycoprotein plays an essential role in the infection process, especialy fusion between virus envelope and cellular endosomic membrane. Recently we reported optimal conditions required for gp64-mediated membrane fusion in pGP64 DNA transfected Spodoptera frugiperda (Sf9) cells (H. J. Kim and J. M. Yang, Jour, Microbiology, 34.7-14). In order to investigate the role of hydrophobicity within the fusion domain of the gp64 glycoprotein for membrane fusion, 13 mutants which have substitution mutation within hydrophobic region I were constructed by PCR-derived site-derected mutagenesis. Each mutated gp64 glycoproteins was transiently expressed by transfecting plasmid DNA into Spodoptera frugiperda (Sf9) cells. Oligomerization of the transisently expressed gp64 glycoproteins was a nalysed by running them on SDS-polyacrylamide gel electrophoresis under non-reducing condition followed by immunoblotting. All of the mutant gp64 glycoproteins expect cysteine-228 were able to form trimers. These results suggest that hydrophobic region I of the gp64 may not be responsible for the oligomerization of the gp64 glycoprotein.

• Applied Microscopy
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• v.28 no.4
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• pp.563-575
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• 1998

To investigate the changes during the differentiation of the cerebral neurons of chick embryo of tne embryogenic day (ED) 7 and 8, the ultrastructural changes in the cerebral neurons, the activity of dehydronases (LDH, MDH and SDH), protein expression profile and adenosine triphosphate concentration were analyzed. In ED 7 chick embryos, relatively large nucleus, centrally located nucleolus, evenly spread chromatin over nucleoplasm, and prominent nuclear envelope were observed. Oval-shaped mitochondria with well-developed cristae were present over entire cytoplasm. In ED 8 chick embryos, evenly spread chromatin over nucleoplasm, and prominent nuclear envelope were observed. In the cytoplasm, well-developed rough endoplasmic reticulum and Golgi complex were observed. In ED 7 chick embryos and ED 8 chick embryos, 31 polypeptide bands and 34 polypeptide bands were observed, respectively. The activities of dehydrogenases were lower in ED 7 chick embryos than in ED 8 chick embryos. LDH activity was 8.16 (ED 7) and 9.28 (ED 8), MDH activity was 7.98 (ED 7) and 10.10 (ED 8), and SDH activity was 5.49 (ED 7) and 7.14 (ED 8) respectively. The ATP concentration remained unchanged over ED 7 and 8.

• The Korean Journal of Zoology
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• v.35 no.2
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• pp.125-135
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• 1992

참개구리의 난자형성 단계에 따른 난황막의 구조적 변화와 막단백질의 변화에 대하여 연구함으로써 난자형성에 따른 난탈막의 기능적 분화의 가능성을 알아보고자 하T:다. 난황막의 구조적 변화는 난자형성에 따른 미세음모의 수와 모양의 변화로 확인되었다. 그 수가 초기에는 적으나 중기에 증가하고 알기에 다시 감소하며, 그 모양도 처음에는 킬고 가늘지만, 나중에는 짧고 굵어진다. 막단백질은 wheat germ agglutinin에 대해서만 특이하게 반응을 보이는 당단백질로서 작은 난모세포(직경 100-800 urn)와 큰 난모세포(직경 1500 urn)에서 다르게 나타난다. 분자량은 106 KD, 60 KD, 순 KD과 같이 어느 단계에서나 공통적으로 나타나는 단백질이 존재하는 반면에 특히 작은 난모세포의 난황막에서만 찾아 볼 수 있는 130 KD, 125 KD, 90 KD, 28 KD, 26KD과 같은 단겐 특이적으로 나타나는 막단백질도 있다. 이와 같은 결과로 미루어 보아 난황막 단백질은 난자형성단계에 따라 변화한다는 사실을 알 수 있다.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.514-517
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• 2001

In order to find potent virus-cell fusion inhibitory components from Korean edible clams, thirteen prepared polysaccharides were introduced to syncytia formation inhibition assay, which is based on the interaction between the HIV-1 envelope protein gp 120/41 and the cellular membutane protein CD4 of T lymphocytes. Among them, Meretrix petechialis showed a potent viruscell fusion inhibitory activity. Fusion index (F1) and percent (%) fusion inhibition of the polysaccharide of this clam were $0.21{\pm}0.02$, and $67.52{\pm}4.09$ at 100781m1, respectively. It exhibited almost equivalent virus-cell fusion inhibitory activity to that of dextran sulfate which was used as a standard control.

• BMB Reports
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• v.35 no.6
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• pp.595-603
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• 2002

A gene that encodes a homologue to baculoviral ODVP-6E/ODV-E56, a baculoviral envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). The ChfuGV odvp-6e/odv-e56 gene was located on an 11-kb BamHI subgenomic fragment using different sets of degenerated primers, which were designed using the results of the protein sequencing of a major 39 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1062 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 353 amino acids with a predicated molecular mass of 38.5 kDa. The amino acid sequence data that was derived from the nucleotide sequence in ChfuGV was compared to those of other baculoviruses. ChfuGV ODVP-6E/ODV-E56, along with othe baculoviral ODVP-6E/ODV-E56 proteins, all contained two putative transmembrane domains at their C-terminus. Several putative N-and O-glycosylation, N-myristoylation, and phosphorylation sites were detected in the ChfuGV ODVP-6E/ODV-E56 protein. A similar pattern was detected when a hydrophobicity-plots comparison was performed on ChfuGV ODVP-6E/ODV-E56 with other baculoviral homologue proteins. At the nucleotide level, a late promoter motif (GTAAG) was located at -14 nt upstream to the start codon of the GhfuGV odvp-6e/odv-e56 gene. a slight variant of the polyadenylation signal, AATAAT, was detected at the position +10 nt that is downstream from the termination signal. A phylogenetic tree for baculoviral ODVP-6E/ODV-E56 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV ODVP-6E/ODV-E56 is most closely related to those of Cydia pomonella granulovirus (CpGV) and Plutella xylostella granulovirus (PxGV).

• Journal of Sericultural and Entomological Science
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• v.33 no.1
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• pp.21-26
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• 1991

Bombyx mori nuclear polyhedrosis virus (BmNPV) was successfully multiplied in the nuclear of BmN4 cells cultured with insect Grace's medium. By electron microscopic observation, the virons had a single nucleocapsid in an envelope. Polyhedral protein synthesis of BmNPV in BmN4 cells was detected at 18 hr p.i. and polyhedral protein was a singlepolypeptide with a M.W of 30 kd. At 48 hr p.i. polyhedra formation was observed by inverted mociroscope and electron microscope. Genome analysis of BmNPV by restriction endonucleases was not revealed the difference between virus produced in vivo and that in vitro.