• Biomolecules & Therapeutics
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• v.17 no.2
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• pp.151-155
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• 2009

Hepatitis C virus (HCV) has genetic diversity like most of RNA viruses. HCV major genotypes are classified into several subtypes which are further divided into quasispecies having, genetically different but closely related variants. The HCV core that is a nucleocapsid protein located at the amino terminus of the viral polyprotein is relatively a conserved protein among the HCV isolates and thus it has been one of plausible targets for anti-HCV drug development. However, different quasispecies of HCV core gene have also been found. In this study, we compared the expression level of core protein between two different quasispecies of HCV genotype 1b. Our data demonstrate that a little differences of amino acid sequence lead to substantial difference of expression level. It might be another important reason of different pathogenesis among HCV infected patients.

• Biomedical Science Letters
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• v.21 no.1
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• pp.9-14
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• 2015

Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

• The Journal of Korean Society of Virology
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• v.26 no.2
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• pp.205-214
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• 1996

한탄바이러스의 내피 단백질 (N)은 이 바이러스에 대한 중요한 항원으로 작용하지만 신증후출혈열 예방과 관련된 작용은 명확히 알려져 있지 않다. 본 연구는 이러한 내피 단백질이 한탄바이러스에 대한 중화 항체를 유도할 수 있는가 하는 관점에서 수행되었다. 한탄바이러스의 내피 단백질을 대장균에서 용해된 형태로 발현하고 이를 단클론 항체를 이용한 면역친화 컬럼으로 분리 정제하였다. 정제된 내피 단백질을 기니픽에 면역하여 항혈청을 얻고 이것의 한탄바이러스에 대한 중화능력을 중화항체 플락 감소법 (plaque reduction neutralization test)을 이용하여 조사한 결과 최고 1:160의 중화능이 있음을 관찰하였다. 이는 한탄바이러스의 내피 단백질이 중화 항체를 유도할 수 있는 epitopes을 가지고 있음을 의리하며 이러한 생각은 본 연구에서 수행한 면역침강법과 N 단백질에 대한 단클론항체를 이용한 면역친화법을 통한 한탄바이러스의 정제 실험 결과에서도 뒷받침되고 있다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.176-183
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• 2016

In this study, disease surveillance was performed to monitor the prevalence of viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) in olive flounder, Paralichthys olivaceus in 2015. The fish samples were collected in March (60 farms), May (55 farms) and July (52 farms) from different farms in Jeju. Reverse transcription polymerase chain reaction (RT-PCR) (VHSV) or PCR (RSIV) results showed that VHSV detected in 2 farms, but RSIV has not been detected in any farms. The sequences of the nucleocapsid protein (N) and glycoprotein (G) gene of the 2 VHSV isolates were successfully sequenced. Phylogenetic analysis was included VHSV isolates reported here together with a representative VHSV isolates available in GenBank. Phylogenetic analysis indicated that most of Korea VHSV isolates were closely related to the Japan and China genotype IVa which is clearly distinct from the North American genotype IVb.

• 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 Fisheries and Marine Sciences Education
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• v.27 no.3
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• pp.879-889
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• 2015

The outbreak of viral diseases caused by viral haemorrhagic septicaemia virus (VHSV) and red seabream iridovirus (RSIV) have been reported in cultured olive flounder, Paralichthys olivaceus. VHSV has been a serious viral disease that infects the olive flounders in South Korea. Clinical signs of VHSV infection are skin darkening, abdominal distension and haemorrhages. Outbreaks of fish iridoviral disease was first reported from red seabream, Pagrus major farms in Japan. Recently, iridovirus infection have occurred frequently from olive flounder farms in South Korea. In this study, disease surveillance was performed to monitor the prevalence of VHSV and RSIV in olive flounder in 2014. The samples were collected from 60 different olive flounder farms in Jeju from April, May, September, November and December in 2014. RT-PCR (VHSV) or PCR (RSIV) results showed that VHSV were detected in 5 farms, but RSIV has not been detected in any farms. The migration of olive flounder was restricted for the quarantine in 5 farms of VHS outbreak. The nucleocapsid protein (N) gene and glycoprotein (G) gene sequences of the 5 Korean VHSV isolates were successfully amplified and sequenced. Phylogenetic analysis was performed using the VHSV sequences reported here together comparison with the nucleotide sequences available from the GenBank database. Phylogenetic analysis indicated that most of Korea VHSV belong to the genotype IVa and closely related to the strains from Japan and China.

• Korean Journal of Veterinary Service
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• v.32 no.1
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• pp.11-18
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• 2009

Porcine epidemic diarrhea virus (PEDV), an enveloped single stranded RNA virus in the family Coronaviridae, causes acute viral enteric disease in piglets. Recently outbreaks of porcine epidemic diarrhea (PED) have been rare in Europe but frequent in Asia. In Korea, the increase of PED prevalence is showing specially in postweaning pigs. The purpose of this study was to investigate nucleotide sequence of nucleocapsid protein gene of PEDV field isolates from postweaning pigs in Korea and get more information about the viruses. A total of 15 postweaing pigs clinically suspected of PEDV infection by severe watery diarrhea and dehydration were used in this study. Viral RNA was extracted from small intestines and stools of the pigs. The N gene was amplified by nested RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. Three PEDVs were isolated from the suspected postweaning pigs. The N gene of three PEDV field isolates consisted of 483 nucleotides. These PEDV field isolates showed nucleotide sequence homology range from 99.6% to 95% with Chinese strains, from 99.8% to 95.2% with Korean strains, from 97.3% to 95.7% with Japanese strains and from 96.5% to 95.7% with Belgium and British strains. The encoded pritein shared range from 98.8% to 95.6% with Chinese strains, from 99.4% to 95% with Korean strains, from 97.5% to 96.3% with Japanese strains, from 95.6% to 95% with Belgium and British strains. By phylogenetic tree analysis based on nucleotide sequence, three PEDV field isolates were clustered into two groups which were Chinese isolate groups and other Korean isolate groups. These results indicated that some of PEDV field isolates prevailing in Korean postweaning pigs may be associated with those of Chinese strains and other Korean strains.

• Journal of Microbiology and Biotechnology
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• v.18 no.10
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• pp.1717-1721
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• 2008

Severe acute respiratory syndrome (SARS) is a life-threatening emerging respiratory disease caused by the coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is highly antigenic and may be a suitable candidate for diagnostic applications. We constructed truncated recombinant N proteins (N1 [1-422 aa], N2 [1-109 aa], and N3 [110-422 aa]) and determined their antigenicity by Western blotting using convalescent SARS serum. The recombinants containing N1 and N3 reacted with convalescent SARS serum in Western blotting. However, the recombinant with N2 did not. In ELISA using N1 or N3 as the antigens, positive results were observed in 10 of to (100%) SARS-CoV-positive human sera. None of 50 healthy sera gave positive results in either assay. These data indicate that the ELISA using N1 or N3 has high sensitivity and specificity. These results suggest that the middle or C-terminal region of the SARS N protein is important for eliciting antibodies against SARS-CoV during the immune response, and ELISA reactions using N1 or N3 may be a valuable tool for SARS diagnosis.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.541-548
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• 1989

The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.107-114
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• 1996

본 연구는 인간면역결핍바이러스의 입자를 비이온성 계면활성제로 처리할 때 바이러스 입자구조에서 분리되어 방출되는 바이러스 구조단백질들의 분포를 sucrose gradient로 분석하여, 바이러스 입자를 구성하는 바이러스 구조단백질과 바이러스입자의 생물리학적 특성을 연구하였다. 바이러스입자들을 0.16% NP40 (Nonidet P-40)으로 처리할 때, 바이러스 capsid 단백질과 바이러스 막 단백질 (membrance protein)들은 다른 바이러스 구성성분들과 잘 분리되었다. 계면활성제처리에서 방출되지 않은 구성 성분들은 matrix 단백질, nucleocapsid 단백질, reverse transcriptase, integrase 및 바이러스 RNA genome로써, 이들은 subviral 구조를 형성한다. 이러한 결과는 상대적으로 다른 바이러스들의 capsid 단백질과 면역 결핍 바이러스의 capsid 단백질 (p24)를 비교할 때, 면역결핍바이러스의 capsid 단백질은 바이러스핵을 형성할 때, capsid 단백질 사이의 결합력이 매우 약한 것으로 추정된다. 또한 바이러스 조절단백질의 하나인 vpr 단백질을 함유하는 바이러스입자를 NP40 처리하여 분석하였을 때, vpr 단백질은 subviral 구조에 존재하는 것으로 나타났다.