• 한국응용곤충학회지
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• 제34권3호
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• pp.218-223
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• 1995

담배와 고추의 주요해충인 담배나방 [Helicoverpa assulta(Guenee)]의 미생물적 방제법을 개발하기 위하여, 아직 국내에서 보고되지 않은 담배나방 핵다각체병 바이러스 담배나방 사충으로 부터 분리하고, 바이러스의 전자현미경 관찰 및 바이러스 DNA의 제한효소 분석 등을 통한 생화학적 특성을 조사하였다. 담배나방 핵다각체병 바이러스의 다각체는 구형에 가까운 20면체로서 크기는 평균 약 1.0$\mu$m 정도이고, 다수의 바이러스 입자가 다각체 단백질에 포매되어 있었다. 포매된 바이러스 입자는 한개의 봉상 nucleocapsid가 하나의 envelope 내에 매립된 형태의 SNPV(single embedded nuclear polyhedrosis virus)로, 형태는 전형적인 봉상으로서 그 크기는 약 $65nm\times300nm$ 였다. 또한 다각체 단백질의 SDS-PAGE 분석 결고, 분자량은 약 31 Kd이었으며, 담배나방 핵다각체병바이러스 DNA를 분리하여 EcoRI. HindIII 등 수종의 제한효소로 처리분석한 결과, 그 genome의 크기는 약 120Kb 정도였다. 본 핵다각체병바이러스는 담배나방 유충에만 병원성을 나타내 기주특이성을 보였다.

• Journal of Microbiology
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• 제34권1호
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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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• 제35권1호
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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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• 제28권4호
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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.

• 한국동물학회지
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• 제35권2호
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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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• 제24권6호
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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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• 제35권6호
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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).

• 한국잠사곤충학회지
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• 제33권1호
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• pp.21-26
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• 1991

Bm 배양세포주(BmN4)에서 BmNPV의 다각체 단백질 합성과 DNA복제에 대한 실험결과는 다음과 같다. 1. BmNPV는 insect Grace's medium에서 배양되고 있는 BmN4 세포에서 NOV나 DNA로 감염시킨 경우 모두 잘 증식되었으며, 도립현미경 관찰 결과 접종 후 48시간이 경과하면서 다각체가 형성되기 시작하였다. 2. 또한 전자현미경으로 핵내 형성된 다각체와 협성중인 nucleocapsid 다발을 관찰하였으며, 바이러스 입자는 SNPV로 존재하였다. 3. Western blot분속에 의한 다각체 단백질의 합성은 접종 후 18시간부터 관찰되었으며, 다각체 단백질의 분자량은 30kd이었다. 4. Wild type BmNPV DNA와 Bm 세포(BmN4)에서 NOV 및 DNA 감염에 의해 형성된 바이러스 DNA간의 제한효소 패턴은 차이가 없었다.

• BMB Reports
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• 제44권9호
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• pp.607-612
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• 2011

Several investigators have shown that CpG-DNA has outstanding effects as a Th1-responsive adjuvant and that its potent adjuvant effects are enhanced by encapsulation with a liposome of proper composition. In this study, we showed that encapsulation with phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE): cholesterol hemisuccinate (CHEMS) complex enhances the immunostimulatory activity of CpG DNA and the binding of CpG-DNA to TLR9. We also examined involvement of myeloid differentiation protein (MyD88) and NF-${\kappa}B$ activation in liposome-encapsulated CpG-DNA-induced IL-8 promoter activation. In this manuscript, the natural phosphodiester bond CpG-DNA encapsulated by DOPE : CHEMS complex is designated as Lipoplex(O). Importantly, we successfully screened B cell epitopes of envelope protein (E protein) of hepatitis C virus (HCV-E) and attachment glycoprotein G of human respiratory syncytial virus (HRSV-G) by immunization with complexes of several peptides and Lipoplex(O) without carriers. Therefore, Lipoplex(O) is potentially applicable as a universal adjuvant for peptide-based epitope screening and antibody production.

• 한국응용곤충학회지
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• 제30권2호
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• pp.144-149
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• 1991

국내에서 분리된 파밤나방 핵다각체병바이러스(Spodoptera exigua nuclear polyhedrosis virus: SeNPV)의 생화학적인 특성을 규명하기 위하여 몇가지 실험을 행하였다. SeNPV는 하나의 envelopeso내에 다수의 nucleocapsid가 존재하는 MNPV(multiple embeded NPV)형태였다. 다각체단백질은 분자량 30kb의 단일 band로 나타났으며, Spodoptera litura NPV와 Bombyx mori NPV의 다각체단백질 항체에 반응하여 뚜렷한 침강선을 형성하였다. 비리온 단백질을 은염색한 결과, 많은 수의 minor band들이 포함된 49개의 band로 나타났으며, 바이러스 DNA를 분리하여 여러종의 제한효소에 의한 대략적인 genome size는 약 110kb 였다.