• Plant Resources
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• 제1권2호
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• pp.92-97
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• 1998

An isolate of barley yellow mosaic virus(BaYMV-HN) obtained from Haenam, Korea was compared with two BaYMV strains. BaYMV-Ⅱ-1 from Japan and BaYMV-G from Germany. The sequence of the 3'-terminal 3817nucleotides[excluding the poly (A) tail] of RNA 1 of BaYMV-HN was determined to start within a long open reading frame coding for a part of the NIa-VPg polymerase(26 amino acids). NIa-Pro polymerase (343 amino acids), NIb polymerase(528 amino acids) and the entire capsid protein(297 amino acids), which is followed by a noncoding region(NCR) of 235 nucelotides. In the partial ORFs, BaYMV-HN shows higher sequence homology with BaYMV-Ⅱ-1(99.5%) than BaYMV-G(92.7%). The 3' non-coding regions of BaYMV-HN(235nt) shows higher nucleotide sequence homology with BaYMV-G(235nt)(99.6%) than BaYMV-Ⅱ-1(231nt)(97.0%). The 3' NIa-Pro protein sequence of BaYMV-HN shows higher amino acid sequence homology with BaYMV-Ⅱ-1(95.0%) than BaYMV-G(93.6%), but, NIb protein sequence of BaYMV-HN shows same all amino acid sequence. The capsid protein sequence of BaYMV-HN(297aa) shows same with BaYMV-Ⅱ-1, and shows higher nucleotide sequence homology with BaYMV-UK (from United Kingdom)(97.3%) than BaYMV-G(96.9%) and G2(96.9%). Difference of capsid protein amino acid were 0-9 between the Japan, United Kingdom and Germany and were 2-6 between all Korean isolates. Many of the amino acid differences are located in the N-terminal regions of the capsid proteins from 1 to 74 amino acid positions.

• Journal of Microbiology
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• 제40권4호
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• pp.313-318
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• 2002

To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported to induce significant humoral and cellular immunity using various animal model systems. In order to produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain reaction, was inserted into yeast expression vector, YEG$\alpha$-HIR525 under the control of GAL10 promoter. The transformation of YEG$\alpha$-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody. The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and simple assay to detect hemagglutihation inhibition activity.

• 한국잠사곤충학회지
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• 제21권2호
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• pp.7-10
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• 1979

1. 연화병 바이러스의 단백질을 SDS-polyacrylamide gel 전기영동으로 분석한 결과 분자량 37,500(FP I), 30,500(FP II) 및 26,500(FP III)에 상당하는 3종류의 polypeptide가 얻어졌으며, 이들의 비율은 FP I이 6.6%, FPII가 25.0% 및 FP III가 68.4%였다. 2. 연화병 바이러스의 단백질을 amino산 분석을 행한 결과, 17종의 amino산이 분리되었으며, 이 amino산 조성은 타곤충 바이러스의 amino산 조성과 유사하였다.

• BMB Reports
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• 제35권6호
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• pp.562-567
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• 2002

In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.482-488
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• 2019

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in pigs. Replicase (Rep) proteins are considered essential for viral replication. Capsid (Cap) protein is the primary immunogenic protein that induces protective immunity. Little is known about comparison on the immunogenicity of PCV2 Rep and Cap fusion protein and Cap protein. In the present study, recombinant baculoviruses expressing the Rep-Cap fusion protein (Bac-Rep-Cap) and the Cap protein (Bac-Cap) of PCV2 were constructed and confirmed with western blot and indirect fluorescence assay. Immunogenicities of the two recombinant proteins were tested in mice. The titers of antibodies were determined with a PCV2-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The $IFN-{\gamma}$ response of immunized mice was measured by ELISA. The mice immunized with the Bac-Rep-Cap and Bac-Cap successfully produced Cap-specific immunoreaction. The mice immunized with the Bac-Cap developed higher PCV2-specific neutralizing antibody titers than mice injected with the Bac-Rep-Cap. $IFN-{\gamma}$ in the Bac-Rep-Cap group was increased compared to those in the Bac-Cap group. Vaccination of mice with the Bac-Rep-Cap showed significantly decreased protective efficacy compared to the Bac-Cap. Our findings will indubitably not only lead to a better understanding of the immunogenicity of PCV2, but also improved vaccines.

• 대한바이러스학회지
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• 제26권2호
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• pp.163-171
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• 1996

The recombinant pseudorabies virus major capsid protein (rMCP) was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. Following evaluation of the immunochemical properties of the rMCP, the immunogenicity of the recombinant subunit protiens were investigated in guinea pig and swine to obtain the preliminary guide line for the subunit vaccine using rMCP and gP50. It was proved that ultrasonication and 30% ammonium sulfate was most efficient to concentrate and purify the protein. The rMCP was safe in mice, guinea pigs and piglets. In guinea pigs, rMCP mixed with various adjuvants induced substantial degree of serum neutralizing antibody titers, but revealed incomplete protectivity against challenge. In swine, the combination of rMCP and gP50 showed the higher serum neutralizing antibody titers and cellular immune responses than rMCP alone. However, the protectivity was lower in comparison with the commercial gI-deleted inactivated vaccine. We expect these results to contribute to characterization of MCP gene of Korean isolate of PRV and to ultilize as preliminary information for prodution and evaluation of PRV recombinant subunit vaccines.

• 한국동물위생학회지
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• 제34권1호
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• pp.1-4
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• 2011

A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.

• Journal of Microbiology
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• 제41권3호
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• pp.239-247
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• 2003

The LRV1-4 capsid protein possesses an endoribonuclease activity that is responsible for the single site-specific cleavage in the 5' untranslated region (UTR) of its own viral RNA genome and the formation of a conserved stem-loop structure (stem-loop IV) in the UTR is essential for the accurate RNA cleavage by the capsid protein. To delineate the nucleotide sequences, which are essential for the correct formation of the stem-loop structure for the accurate RNA cleavage by the viral capsid protein, a wildtype minimal RNA transcript (RNA 5' 249-342) and several synthetic RNA transcripts encoding point-mutations in the stem-loop region were generated in an in vitro transcription system, and used as substrates for the RNA cleavage assay and RNase mapping studies. When the RNA 5' 249-342 transcript was subjected to RNase T1 and A mapping studies, the results showed that the predicted RNA secondary structure in the stem-loop region using FOLD analysis only existed in the presence of Mg$\^$2＋/ ions, suggesting that the metal ion stabilizes the stem-loop structure of the substrate RNA in solution. When point-mutated RNA substrates were used in the RNA cleavage assay and RNase T1 mapping study, the specific nucleotide sequences in the stem-loop region were not required for the accurate RNA cleavage by the viral capsid protein, but the formation of a stem-loop like structure in a region (nucleotides from 267 to 287) stabilized by Mg$\^$2＋/ ions was critical for the accurate RNA cleavage. The RNase T1 mapping and EMSA studies revealed that the Ca$\^$2＋/ and Mn$\^$2＋/ ions, among the reagents tested, could change the mobility of the substrate RNA 5' 249-342 on a gel similarly to that of Mg$\^$2＋/ ions, but only Ca$\^$2＋/ ions identically showed the stabilizing effect of Mg$\^$2＋/ ions on the stem-loop structure, suggesting that binding of the metal ions (Mg$\^$2＋/ or Ca$\^$2＋/) onto the RNA substrate in solution causes change and stabilization of the RNA stem-loop structure, and only the substrate RNA with a rigid stem-loop structure in the essential region can be accurately cleaved by the LRV1-4 viral capsid protein.

• Fisheries and Aquatic Sciences
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• 제9권4호
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• pp.146-152
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• 2006

Systemic infections of maricultured fishes by Megalocytivirus species have occurred over a broad area in South Korea, causing extensive economic loss. We developed digoxigenin-labeled nucleic acid probes against the 230-bp ATPase and 311-bp major capsid protein (MCP) of rock bream Oplegnathus fasciatus iridovirus (RBIV) using polymerase chain reaction, and an in situ hybridization (ISH) method to detect Megalocytivirus in formalin-fixed tissues of mariculture species (rock bream, sea bass, and olive flounder). ISH-positive cells were abundant in the hematopoietic and connective tissues of various organs, while brain tissue showed little or no signal. The ISH procedure can become an important diagnostic tool in complement with histopathological methods, and advances epidemiological studies on the origin and distribution of Megalocytivirus in mariculture.

• 한국어병학회지
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• 제32권2호
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• pp.49-57
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• 2019

본 연구에서는 2012년부터 2018년 1월까지 국내 어류에서 참돔이리도바이러스병 (red sea bream iridoviral disease; RSIVD) 으로 확정 진단된 주요 분리주에 대하여 major capsid protein (MCP) 유전자의 염기서열을 바탕으로 Megalocytivirus의 유전적 관계를 명확히 했다. 또한, Megalocytivirus와 숙주 세포간 상호작용에 영향을 줄 수 있는 특이 유전자 2종, Vascular endothelial growth factor (VEGF) 및 Histidine triad motif인 HIT-like 단백질 (HIT)에 대한 염기서열 분석을 통해 유전적 상관관계를 고찰하였다. 국내 어류에서 분리된 39개의 megalocytiviruses는 2가지 유전형인 red sea bream iridovirus (RSIV)형과 turbot reddish body iridovirus (TRBIV)형으로 분류되었으며, RSIV형의 megalocytiviruses는 다시 유전아형인 RSIV-subgroup 1형과 2형으로 분류되었다. 그리고 VEGF 유전자 염기서열 분석 결과에서는 RSIV형의 바이러스에서 변이가 발생되었음을 확인할 수 있었으며, HIT-like 단백질 유전자는 RSIV형의 Megalocytivi rus에서만 발견되는 것을 알 수 있었다.