• Korean Journal of Veterinary Research
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• v.35 no.4
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• pp.743-752
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• 1995

The study was carried out to characterize the properties of Korean isolates of EHV from aborted fetuses and determine envelope protein profiles. The results obtained were summarized as follows; 1. Two strains of EHV was isolated from 2 liver samples among 10 aborted fetuses from which the virus isolation was attempted. 2. Morphological and some enzymatic properties of the Korean isolates of EHV which was designated as $LC_1$ and $LC_2$ was identical to those of a reference strain of Australia-N of EHV-1. The Korean isolates of EHV could be propagated on ED cell culture and they formed typical plaques 1 to 2 days after infection in the ED cells from which typical cuboidal particles of 150~170 nm diameter herpesvirus were observed. The virus could be detected specifically from neucleus and cytoplasm of infected cells by flourescent antibody technique using FITC labelled anti-Aust IV(EHV-1) antiserum. The Korean isolates, $LC_1$ and $LC_2$ were specifically neutralized by anti Aust IV antiserum and reacted positively to CELISA. 3. The structural polypeptides of purified enveloped virions of $LC_1$ and $LC_2$ isolates of EHV were determined by SDS-polyacrylamide gel electrophoresis to identify the envelope glycoproteins. $LC_1$ and $LC_2$ strains revealed 14 glycoproteins ranging in molecular weight from 190 kD to 31 kD while 17 structural proteins of Aust IV(EHV-1), of which 14 were identical to those of $LC_1$ and $LC_2$, were identified. Upon immunoblotting by rabbit antiserum against EHV isolates and EHV-1(Aust IV), 4 immunogenic proteins of $LC_1$ and $LC_2$ were 135 kD, 88 kD, 64 kD and 59 kD, of which 135 kD, 88 kD and 64 kD proteins were also found in Aust IV(EHV-1).

• The Plant Pathology Journal
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• v.16 no.5
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• pp.269-279
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• 2000

Alfalfa mosaic alfamoviruses(AIMV) were isolated from infected potato (Solanum tuberosum) and azuki bean (Paseolus angularis) in Korea. Two AIMV isolated from potatoes were named as strain KR (AIMV-KR1 and KR2) and AIMV isolated from azuki bean was named as strain Az (AIMV-Az). Each isolated AIMV strain was characterized by using their host ranges, symptom developments, serological relations and nucleotide sequence analysis of coat protein (CP) gene. Strains KR1, KR2, and Az were readily transmitted to 20 of 22 inoculated plant species including bean, cowpea, tomato, tobacco, and potato. AIMV-KR1 and KR2 produced the typical symptoms like chlorotic or necrotic spots in Chenopodium quinoa and Solanum tuberosum cv. Superior. AIMV-Az caused bright yellow mosaic symptom and leaf malformation in Nicotiana glauca, which were different from the common mosaic symptom caused by AIMV-KR1 and KR2. Electron microscope observation of purified virus showed bacilliform virions containing a single-stranded plus-strand RNAs of 3.6, 2.6, 2.0 and 0.9 kbp in length, respectively, similar in size and appearance to those of Alfamovirus. In SDS-PAGE, the coat protein of the two viruses formed a consistent band that estimated to be about 24kDa. The CP genes of the AIMV strains, KR1, KR2, and Az have been amplified by RT-PCR using the specific primers designed to amplify CP gene from viral RNA-3, cloned and sequenced. Computer aided analysis of the amplified cDNA fragment sequence revealed the presence of a single open reading frame capable of encoding 221 amino acids. The nucleotide and peptide sequence of viral CP gene showed that strain KR1, KR2, and Az shared highest nucleotide sequence identities with AIMV strain 425-M at 97.7%, 98.2%, and 97.2%, respectively. CP gene sequences of two strains were almost identical compared with each other. Altogether, physical, serological, biological and molecular properties of the purified virus.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.121-129
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• 1996

Multiplication of Herpes simplex virus type-1 was observed by electronmicroscopy, a gene library of the genome was constructed and thymidine kinase gene was cloned. Vero cells infected with the virus were lysed 48 h p.j. and multinucleated giant cells were observed approximately at 72 h p.i. The nucleocapsids were observed in nuclei and cytoplasm, and the assembled nucleocapsids were budded out through the vacuole and cytoplasmic membranes, and then virions were released from the cells. HSV-1 genome DNA was digested with BamHI and BglII enzymes and then the gene library of the genome fragments were constructed. The BamHI cleaved the genome DNA into twenty-seven fragments in the range of 1.1 - 14 kb, and BglII cleaved the genome DNA into sixteen fragments in the range of $4.5{\sim}20.1\;kb$. The pHLA-12 and pHLB-4 recombinant plasmids were contained TK gene by Southern blot analysis. The molecular sizes of the fragments which contained the TK gene were 3.74 in pHLA-12 and 6.41kb in pHLB-4 recombinant plasmid, respectively.

• Journal of fish pathology
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• v.11 no.2
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• pp.129-136
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• 1998

In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

• The Plant Pathology Journal
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• v.19 no.2
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• pp.117-122
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• 2003

Six isolates of Turnip mosaic Potyvirus (TuMV) namely, TuMV-CA7 from oriental cabbage, TuMV-TU and TuMV-TU2 from turnip, TuMV-RA from rape, TUMV-ST from stock, and TuMV-R9 from radish, and Ribgrass mosaic Tobamovirus (RMV-FG22) from oriental cabbage were isolated. Three kinds of characteristics of the six TuMV isolates were sorted by bioassay: TuMV-CA7 and TuMV-TU isolates infected mostly oriental cabbages; TuMV-ST, TuMV-TU2, and TuMV-R9 infected radishes; and TuMV-RA infected both oriental cabbages and radishes. Mixed infections of crucifers were RMV-FG22+TuMV-CA7, RMV-FG22+TuMV-TU, RMV-FG22+TuMV-RA, RMV-FG22+TuMV-ST, RMV-FG22 +TuMV-TU2 and RMV-FG22+TuMV-R9. Crops used were 'Tambok' cultivar resistant to TuMV, 'SSD63' susceptible inbred line of oriental cabbage, pure line of leaf mustard and 'Daeburyungyeorum' cultivar of radish. New specific ultrastructures of nonagon-like ring (NLR) and spiral aggregates (SA) by mixed infection with TuMV and RMV were formed in cells of crucifer plants. The NLR was made by a TuMV surrounded loosely by nine RMV particles, and the SA was formed spirally by full mixed of two virus particles. The SA had some NLR in its center, which was observed from cross sectioned SA. Host plants with specific ultrastructures expressed synergistic symptoms. Specific ultrastructures of NLR and SA were formed in combinations of RMV-FG22 and in TuMV-CA7, TuMV-TU, or TuMV-RA that could infect oriental cabbages. How-ever, no specific ultrastructures and mixing of the two virions in the same cell were observed in combinations of RMV-FG22, and TuMV-57, TuMV-TU2, or TuMV-R9 isolates haying virulence in radishes.

• Research in Plant Disease
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• v.13 no.2
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• pp.75-81
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• 2007

The incidence of virus disease on peppers was investigated at the 52 areas in the whole country in 2002, 2004 to 2006. Among the six viruses, Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV) and Tomato spotted wilt virus (TSWV), infecting peppers in Korea, the incidence of CMV, PepMoV, PMMoV and BBWV2 was 29.4%, 13.6%, 14.3%, 25.6%, orderly. TMGMV and TSWV had the same low infection rate of 1.0%. The infection rate of CMV was higher as 53.3% and 34.2% in 2002 and 2004, but it was decreased to 18.2% and 11.9% in 2005 and 2006, respectively. The infection rate of BBWV2 was lower as 1.3% in 2002 and 1.8% in 2004, but it was increased abruptly to 41.3% in 2005 and 58.2% in 2006. For the types of mixed infections of pepper viruses, the incidence of CMV+PepMoV was 62.6% in 2002 and 50.0% in 2004, and that of CMV+BBWV2 was increased suddenly from 33.3% in 2005 to 83.2% in 2006. The triplex infection of CMV+BBWV2+PepMoV was 6.4% in average. CMV caused severe mosaic and BBWV2 induced ring spots, and the two mixed virions caused chlorosis on the leaves of red peppers. TSWV induced the typical symptoms of multiple ring spots on the leaves and fruits of red peppers.

• Journal of Microbiology and Biotechnology
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• v.28 no.8
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• pp.1376-1383
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• 2018

The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity ($K_D$) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.

• Korean Journal of Microbiology
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• v.45 no.3
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• pp.246-250
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• 2009

Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.