• Korean Journal of Veterinary Service
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• v.25 no.2
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• pp.153-174
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• 2002

파라믹소바이러스과 (Parnxouiridae) 바이러스는 외피당단백질의 생물학적 특성에 있어서 Orthomyxoviridae와, 분절되지 않은(nonseg-mented)게놈과 그 발현의 구성 측면에서 Rhbdoviridae와 유사성을 갖는 바이러스로서 외피로 둘러 싸여 있는 마이너스쇄(negative strand) RNA 바이러스이다. 마이너스쇄 RNA 바이러스의 게놈 RNA는 2가지 기능으로 작용한 다. 즉, 첫째는 mRNA 합성을 위한 주형(鑄型, template)으로 작용하고, 둘째는 플러스쇄 (anti-genome(+) strand)의 합성을 위한 주형으로서 작용한다. 마이너스쇄 RNA바이러스는 그들 자신의 RNA전사효소를 코드화 하여 저장하지만, mRNA는 감염세포 내에서 게놈이 노출된 후에만 합성된다. 바이러스 복제는 mRNA의 합성후 일어나며 연속된 바이러스단백질의 합성을 필요로 한다. 새로 합성된 플러스쇄(antigenome(+)strand)는 마이너스쇄 게놈 RNA의 복제를 도모하기 위한 주형(鑄型)으로서 작용한다.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.8
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• pp.5412-5418
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• 2014

The red seabream iridovirus (RSIV), which belongs to the iridoviridae, causes infectious fish diseases in many Asian countries, leading to considerable economic losses to the aquaculture industry. Using the yeast surface display (YSD) technique, a new experimental system was recently developed for the detection and identification of a variety of marine viruses. In this study, a coat protein gene of RSIV was synthesized based on the nucleotide sequence database and subcloned into the yeast expression vector, pCTCON2. The expression of viral coat proteins in the yeast strain, EBY100, was detected by flow cytometry and Western blot analysis. Finally, they were isolated from the yeast surface through a treatment with ${\beta}$-mercaptoethanol. The data suggests that the YSD system can be a useful method for acquiring coating proteins of marine viruses.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.140-147
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• 2010

Japanese encephalitis virus (JEV), a member of the mosquito-borne flaviviruses, causes epidemics of viral encephalitis in the Southeastern Asia. JEV is a small enveloped virus with a positive-sense RNA genome; the infectious virion consists of three structural proteins, namely capsid, membrane (M; a mature form of its prM precursor), and envelope proteins. Here, we investigated a role of the charged residues found at the N-terminus of the JEV M protein in virus production. Using an infectious JEV cDNA, we generated two mutant cDNAs, Mm1 and Mm2, by charged-to-alanine substitution for $E^9$ and $K^{15}K^{16}E^{17}$ residues of the M protein, respectively. By transfection of wild-type or each of the two mutant RNAs transcribed from the corresponding cDNAs, we found that Mm2, but not Mm1, had a ~3-log decrease in virus production, even though a comparable amount of all three structural proteins were produced in transfected cells. Interestingly, the prM protein expressed in Mm2 RNA-transfected cells was not recognized by the polyclonal antiserum raised against the N-terminal 44 amino acids of the wild type M protein, but reacted to the antiserum raised against the corresponding region of the mutant Mm2. Our results indicate that three charged residues ($K^{15}K^{16}E^{17}$) in JEV M protein play a role in virus production. Two polyclonal antisera specifically recognizing the wild-type or Mm2 version of the M protein would provide a useful reagent for the functional study of this protein in the virus life cycle.

• Korean Journal Plant Pathology
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• v.12 no.2
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• pp.182-186
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• 1996

국내에서 재배되고 있는 고추(Capsicum annuum L.)로부터 분리된 TMV pepper 계통을 density gradient centrifugation을 이용하여 순화하였다. 이로부터 바이러스의 total RNA를 분리하였고 RT-PCR에 의하여 TMV pepper 계통의 외피단백질 cDNA를 합성, 증폭하였으며 이를 pBluescript II SK- 벡터에 재조합하였다. 본 실험에서 바이러스 외피단백질과 3` non-coding region을 포함하는 재조합 클론 p1561과 p1562로부터 염기서열을 분석하였고 그 결과로 477 염기의 외피단백질 유전자를 포함하는 691 염기가 합성되었음을 확인하였으며 이것과 TMV common 계통으로부터 합성된 외피단백질 cDNA와의 최대 유사도는 69%였다. 또한 유추된 아미노산 서열에서 이들 두 계통간의 최대 유사도는 81%였다.

• Korean journal of applied entomology
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• v.30 no.2
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• pp.144-149
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• 1991

Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

• 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.

• Korean journal of applied entomology
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• v.30 no.3
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• pp.219-226
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• 1991

A cytoplasmic polyhedrosis virus isolated from the oriental tobacco budworm, Heliothis assulta (HaCPV), was studied on morphology of the polyhedron and virus particles, analysis of viral protein and nucleic acid, and bioassay of the HaCPV to determine the feasibility of application as a microbial control agent. The shape of polyhedron was hexagonal ranging 0.5-3.7 ${\mu}m$ and the virus particles were icosahedral outline measured 55 nm in diameter. Polyhedral protein was composed of a major polypetide of 24.3 Kd and 5 minor components and virus particle had seven polypeptides ranging in 28.0 Kd-133. 6 Kd by the SDS-P AGE. The genome of virus was segmented with 10 double stranded RNA in the total mol. wt. of 18.08 Md ranging in 0.65 Md -2.79 Md. The $LC_{50}$ values of the HaCPV to the 3rd instar of H. assulta larvae were calculated to $2.895{\times}10^5PIBs/ml$. The $LT_{50}$ values in the concentration of $5.0{\times}10^{6}PIBs/ml$ was 16.4 days.

• Journal of Sericultural and Entomological Science
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• v.38 no.2
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• pp.144-149
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• 1996

To develope the baculovirus expression vector system (BEVS) using Spodoptera exigua nuclear polyhedrosis virus (SeNPV), we characterized the polyhedrin of SeNPV. The SeNPV polyhedra was irregular and composed of the major protein molecular weight of 30 kDa determined by electronmicroscopy and SDS-AGE analysis, respectively. The nucleotid suquences of 876 bases including the coding region of polyhedrin gene was determined and it was revealed that the polyhedrin gene is located within Xho I 3.0Kb and Nco I 6.0 Kb by Southern blot analysis, respectively. Also, the Xho I 3.0 Kb and the Nco I 6.0 Kb fragments were cloned and restriction enzyme map of these clones were determined.

• Research in Plant Disease
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• v.12 no.1
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• pp.25-27
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• 2006

Rice dwarf phytoreovirus (RDV), a member of the family Reoviridae has a genome composed of 12 segmented dsRNAs designated as 51 to 512 with an increasing order of mobility in polyacrylamide gel electrophoresis (PAGE). RDV encode 12 structural and non-structural proteins, $P1{\sim}P12$ which are encoded by the $S1{\sim}S12$ segments of the dsRNA genome, respectively. In this experiment, we confirmed in situ localization of RDV particles and P12 in cytoplasm of infected rice plant. We observed specific reaction of the gold particles using virus particle and P12 protein specific antiserum with protein A-gold immunolabelling in electron microscope. It was observed that gold particles specifically react to virus particles in cytoplasm in case using the antiserum for virus particles. In the case of antiserum for P12 protein, gold particles sporadically existing on cytoplasm without existing in organelle of cytoplasm specifically. As this result, RDV P12 protein encoded by S12 located in cytoplasm.

• Journal of Life Science
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• v.26 no.12
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• pp.1470-1476
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• 2016

The viral hemorrhagic septicemia virus (VHSV), which belongs to the Novirhabdovirus genus of the Rhabdoviridae family, is a viral pathogen that causes severe losses in the olive flounder farming industry. Among six encoding VHSV proteins, the non-virion (NV) protein has been shown to have an impact on virulence. In our previous studies, transcriptomics microarray analysis by using VHSV-infected olive flounder showed that VHSV infection significantly down-regulated the mRNA expression of glycolytic enzymes. In addition, VHSV NV protein variants decreased the intracellular ATP level. Based on these results, we have tried to examine the effect of VHSV NV protein on glycolytic enzyme glucokinase expression, which phosphorylates glucose to glucose 6-phosphate. Our results indicated that the NV protein significantly decreased the mRNA expression of glucokinase in olive flounder HINAE cells. Furthermore, the NV protein played a negative role in the promoter activation of glucokinase. Furthermore, glucose uptake was effectively inhibited by VHSV infection and NV protein expression in olive flounder HINAE cells. These results suggest that the VHSV NV protein negatively regulates glycolytic enzyme expression by a transcription level and eventually leads to gradual morbidity of olive flounder through cellular energy deprivation. The present results may be useful for the prevention and diagnosis of VHSV infection in olive flounder.