• 한국동물위생학회지
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• 제32권1호
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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.

• 수산해양교육연구
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• 제27권3호
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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.

• The Plant Pathology Journal
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• 제20권4호
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• pp.297-301
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• 2004

A Tomato spotted wilt virus (TSWV-KP) was isolated from Paprika (Capsicum annuum var. grossum) showing necrosis spot on the leaves and malformation of the fruit in Yesan, Korea. The virus infected Chenopodium amaranticolor, C. quinoa, Petunia hybrida, Nicotiana glutunosa, Gomphrena globosa, and Physalis floridana. Ten plants including tomato were observed to have systemic TWSV-KP infection. The virus produced necrosis or necrotic ring spots on the inoculated leaves and mosaic, vein necrosis or death on the upper leaves of Datura stramonium, N. clevarandii, N. rustica, and N.tabacum cvs. Thin sections of the infected leaf tissue contained spherical to oval particles, a characteristic of a Tospovirus. The virion contained three molecules of genomic RNAs, which were approximately 9.0, 4.9 and 3.0 kb. The nucleocapsid (N) protein of the purified virion migrated as a single band with molecular weight of about 29 kDa in SDS-PAGE. The N gene of TSWV-KP showed 96.5-97.2% and 97.7-98.5% identities to the three different TSWV isolates of Genbank Database at the nucleotide and amino acid, respectively.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2003년도 추계학술대회초록집
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• pp.41-41
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• 2003

White spot syndrome virus (WSSV) is the causative agent of a disease that has led to severe mortalities of cultured shrimps in Korea and many other countries. Since 1993, massive mortalities due to the viral infection have also occurred in the penaeid shrimps cultured in Korea. WSSV is a large, circular, double stranded (ds) DNA virus and an enveloped, ellipsoid virus with a rod-shaped nucleocapsid with flat ends. In order to identify the characteristics of this Korean isolate of WSSV, the genes for four virion proteins, VP15, VP19, VP28 and VP35 were cloned and their sequences were compared with the available pool of WSSV gene sequences in the GenBank/EMBL databases. From these comparisons, we confirm the occurrence of WSSV in Korea and deduce that, VP15, VP28 and VP35 genes are identically conserved among the Korean isolate and geographically different foreign isolates, but VP19 amino acid sequences of the Korean WSSV isolates changed valine of the foreign isolates into aspartate. (omitted)

• 대한바이러스학회지
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• 제26권1호
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• pp.121-129
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• 1996

Herpes simplex virus type-1의 vero 세포에서의 증식기작을 규명하기 위해 전자현미경으로 관찰하였고, 유전학적 특성을 규명하기 위해 유전자도서관을 작성하였고, thymidine kinase (TK) 유전자를 클로닝을 하였다. 감염 48시간 후 많은 수의 바이러스 nucleocapsid가 핵뿐만 아니라 세포질에서 관찰되었다. 바이러스는 세포에 감염된 후 핵내에서 복제증식한 후 세포질내로 이동하였으며, 이때 핵막을 통과하면서 외투막을 갖고 세포질로 이동하여 세포밖으로 나가는 것을 관찰할 수 있었으며, 또한 일부 nucleocapsid는 세포막을 출아하여 비리온으로 출아되었다. HSV-1의 DNA를 BamHI과 BglII 제한효소로 각각 절단하여 DNA의 절단 양상을 조사하였다. BamHI에 의해 절단된 단편은 27개 이었고, 그들 분자량의 범위는 1.1 - 14 kb이었으며, BglII에 의해 절단된 단편은 16개이었고, 분자량의 범위는 4.5 - 20.1 kb이었다. Southern blot 방법으로 TK 유전자를 포함하고 있는 단편을 확인하였는데, pHLA-12와 pHLB-14클론에 포함되어 있었고 각 단편의 분자량은 3.74와 6.41 kb이었다.

• 한국미생물·생명공학회지
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• 제37권3호
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• pp.287-292
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• 2009

돼지생식기호흡기증후군 바이러스를 구성하고 있는 뉴클레오캡시드(N) 단백질은 다양한 기능을 가지고 있는 basic 단백질로써 또한 아직까지 밝혀지지 않은 역할을 하는 serine 인산화 단백질로 알려져 있다. 먼저 바이러스가 복제되는 동안 뉴클레오캡시드 단백질 인산화가 어떤 생물학적 역할을 하는지에 대한 이해를 하기 위하여 mutagenesis 방법으로 단백질 내 모든 serine 잔기들을 alanine으로 대체하여 변이 뉴클레오캡시드 단백질을 구축하였다. 이 재조합 뉴클레오캡시드 단백질은 비인산화 단백질로 확인되었고 이는 뉴클레오캡시드 단백질 인산화에 serine 잔기들이 중요한 역할을 한다는 것을 증명하였다. 돼지 생식기호흡기증후군 바이러스 뉴클레오캡시드 단백질은 세포핵 내 이동과 N-N dimer 형성 등의 특이적인 생물학적 특성들을 보유하고 있으며 이들 각각은 바이러스 감염 시 중요한 역할들을 하는 것으로 알려져 있다. 따라서 본 연구에서는 이 두 가지 뉴클레오캡시드 단백질의 특성들이 인산화 여부에 의해 조절되는지 살펴보았다. 하지만 본 연구의 결과들은 비인산화된 뉴클레오캡시드 단백질이 여전히 transfection된 세포의 핵 또는 핵인에서 발현되었고 더욱이 뉴클레오캡시드 자신과 dimer 형성을 할 수 있었다는 것을 보여주었다. 결론적으로 돼지 생식기호흡기증후군 바이러스 뉴클레오캡시드 단백질의 세포핵 내 수송 및 oligomerization 특성들은 인산화 비의존성으로 조절되는 것으로 보여 진다. 아마도 이 인산화 작용은 뉴클레오캡시드 단백질의 RNA-binding 특성등과 같은 다른 수준의 조절과 관련이 있는 것으로 추측되어 진다.

• 대한수의학회지
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• 제43권2호
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• pp.219-230
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• 2003

Porcine epidemic diarrhea virus(PED), a member of Coronaviridea, is the etiological agent of enteropathogenic diarrhea in swine. The purpose of this study was to investigate genetic characteristic of PEDV isolated in Korea. Nucleocapsid(N) gene and membrane (M) gene of recent Korean PEDV strains isolated in 2001 were amplified, cloned, sequenced and analyzed. N gene of seven Korean PEDV field isolates bad 94.5% to 99.4% nucleotide and 92.4% to 99.4% amino acid sequence homology each other. Nucleotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 95.1% to 98.0% nucleotide and 93.5% to 97.6% amino acid sequence homology. By phylogenetic tree analysis on based nucleotide sequences, PEDVs were clustered into four groups. By phylogenetic tree analysis based on amino acid sequences. PEDVs were clustered into five groups. M gene of our Korean PEDV field isolates had 99.6% to 100% nucleotide and 98.7% to 100% amino acid sequence homology each other. Nuclotide and amino acid sequences of Korean field PEDVs were different from published foreign PEDVs, showing 98.5% to 98.8% nucleotide and 97.3% to 97.8% amino acid sequence homology. By phylogenetic tree analysis based on nucleotide and amino acid sequences, PEDVs were clustered into two groups which were Korean PEDV isolate group and foreign PEDV isolate group.

• 한국수산과학회지
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• 제49권2호
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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.

• The Plant Pathology Journal
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• 제34권6호
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• pp.514-531
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• 2018

Tomato spotted wilt virus (TSWV; Genus Orthotospovirus: Family Tospoviridae) is one of the most destructive viruses affecting a wide range of horticultural crops on a worldwide basis. In 2015 and 2016, 171 leaf and fruit samples from tomato (Solanum lycopersicum) plants with viral symptoms were collected from the fields in various regions of Iran. ELISA test revealed that the samples were infected by TSWV. The results of RT-PCR showed that the expected DNA fragments of about 819 bp in length were amplified using a pair of universal primer corresponding to the RNA polymerase gene and DNA fragments of ca 777 bp and 724 bp in length were amplified using specific primers that have been designed based on the nucleocapsid (N) and non-structural (NSs) genes, respectively. The amplified fragments were cloned into pTG19-T and sequenced. Sequence comparisons with those available in the GenBank showed that the sequences belong to TSWV. The high nucleotide identity and similarities of new sequences based on the L, N, and NSs genes showed that minor evolutionary differences exist amongst the isolates. The phylogenetic tree grouped all isolates six clades based on N and NSs genes. Phylogenetic analysis showed that the Iranian isolates were composed a new distinct clade based on a part of polymerase, N and NSs genes. To our knowledge, this is the first detailed study on molecular characterization and genetic diversity of TSWV isolates from tomato in Iran that could be known as new clade of TSWV isolates.

• 대한수의학회지
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• 제52권1호
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• pp.9-18
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• 2012

Rabies is a major zoonotic disease that causes approximately 55,000 human deaths worldwide on an annual basis. The nucleocapsid protein and glycoprotein genes of the Korean rabies virus (RABV) have been subjected to molecular and phylogenetic analyses. Although the phosphoprotein (P) has several important functions in viral infection and pathogenicity, the genetic characterizations of the P of Korean RABV isolates have not yet been established. In the present study, we conducted genetic analyses of P genes of 24 RABV isolates circulating in the Republic of Korea (hereafter, Korea) from 2008 to 2011. This study revealed that the P genes of Korean RABVs are genetically similar to those of RABV strains of lyssavirus genotype I including V739 (dogs, Korea), NNV-RAB-H (humans, India), NeiMeng925 (raccoon dogs, China), and RU9.RD (raccoon dogs, Russia). Among Korean isolates, the RABV P genes showed low variability in the variable domains among Korean isolates; they had specific consensus sequences and amino acid substitutions capable of identifying geographic characteristics and retained specific sequences thought to be important for viral function. These results provide important genetic characteristics and epidemiological information pertaining to the P gene of the Korean RABV.