• 제목/요약/키워드: Major capsid protein (MCP)

검색결과 16건 처리시간 0.031초

Baculovirus Vector System에 의해 발현된 재조합 Pseudorabies Virus Major Capsid Protein의 면역원성 (Immunogenicity of the Recombinant Pseudorabies Virus Major Capsid Protein Expressed by Baculovirus Vector System)

  • 전무형;안동준;장경수;조용성;박종현;송재영;현방훈;안수환
    • 대한바이러스학회지
    • /
    • 제26권2호
    • /
    • pp.163-171
    • /
    • 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.

  • PDF

The serodiagnosis of a lymphocystis disease virus infection using an antibody raised against a recombinant major capsid protein

  • Seo, Ja-Young;Kang, Bong-Jo;Oh, Hyoung-Jong;Lee, Jae-Il;Kim, Tae-Jung
    • 한국어병학회지
    • /
    • 제21권3호
    • /
    • pp.175-180
    • /
    • 2008
  • Lymphocystis is a viral disease of fish primarily in marine and brackishwaters. Here we report the cloning, expression, and the serological applications of the lymphocystis disease virus (LCDV) major capsid protein (MCP). The MCP gene was amplified by PCR from the genomic DNA of LCDV isolated from Schlegel's black rockfish, Sebastes schlegeli, and expressed in E. coli. Mouse antisera raised against the purified recombinant MCP (rMCP) reacted with the viral MCP in an immunofluorescence assay, indicating that this rMCP would be useful for serological studies of field samples.

Pseudorabies Virus의 Major Capsid Protein 유전자의 클론닝과 Baculovirus Vector System에 의한 발현 (Cloning of Major Capsid Protein Gene of Pseudorabies Virus and Expression by Baculovirus Vector System)

  • 안동준;전무형;송재영;박종현;현방훈;장경수;안수환
    • 대한바이러스학회지
    • /
    • 제26권2호
    • /
    • pp.151-162
    • /
    • 1996
  • Pseudorabies is caused by Pseudorabies virus (PRV: Aujeszky's disease virus) of Herpesviridae that is characterized by 100 to 150nm in size with a linear double-stranded DNA molecule with of approximately $90{\times}10^6Da$. This disease affects most of domestic animals such as swine, cattle, dog, sheep, cat, chicken, etc. causing high mortality and economic losses. In swine, young piglets show high mortality and pregnant sows, reproductive failures. However the adult swine reveals no clinical signs in general. But they become a carrier state and play an important role for propagation of the disease. In this study, the nucleotide sequence of major casid protein gene of PRV, Yangsan strain isolated from the diseased swine in Korea was analyzed, and the recombinant MCP was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. As result, in BamHI digestion, MCP gene locus of PRV YS strain showed different from that of Indiana S strain. The patterns of enzyme mapping were also found to be unidentical each other. The sequence of the MCP gene partially analyzed showed 98.09% identity to Indiana S strain. The expression of MCP in Sf-9 cell cotransfected by pVLMCP-44 baculovirus expression vector was characterized by Southern blot hybridization, immunofluoresent and immunocytochemical tests, SDS-PAGE and Western blotting. The rMCP with M.W. 142kDa was most effectively expressed in Sf-9 cells at the 3-4th days post inoculation of the recombinant baculovirus by 2 moi.

  • PDF

국내 어류에서 분리된 Megalocytivirus의 유전형 분류 및 상관관계 분석 (Genetic relatedness of Megalocytivirus from diseased fishes in Korea)

  • 이은선;조미영;민은영;정승희;김광일
    • 한국어병학회지
    • /
    • 제32권2호
    • /
    • pp.49-57
    • /
    • 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.

Comparison of detective ranavirus with major capsid protein gene from infected frogs (Pelophylax nigromaculatus and Lithobates catesbeianus) in South Korea

  • Jongsun, Kim;Nam-Ho, Roh;Jaejin, Park;Daesik, Park
    • Journal of Ecology and Environment
    • /
    • 제46권4호
    • /
    • pp.276-281
    • /
    • 2022
  • Ranaviruses are a primary cause of amphibian extinctions. More consistent ranavirus-infection reports and genetic characterizations of identified viruses are urgently needed, particularly from Asian countries. The objectives of this study were to obtain the partial major capsid protein (MCP) gene sequences (506 bp) of the ranavirus responsible for infecting frogs in South Korea, as our previous research had confirmed using qPCR, and to evaluate their genetic relationships with other previously reported ranavirus sequences. Three different ranavirus MCP sequences were obtained from Pelophylax nigromaculatus and Lithobates catesbeianus. All six different types of MCP sequence from the ranavirus identified in South Korea to date belonged to the Frog virus 3 (FV3)-like virus group in the genus Ranavirus. To better understand the origin and spread of ranaviruses in South Korea, further infection reports and full genome analyses of the identified ranaviruses are needed.

In situ Hybridization of a Megalocytivirus Using Nucleic Acid Probes against ATPase and the Major Capsid Protein of Rock Bream Iridovirus

  • Lee, Nam-Sil;Do, Jeong-Wan;Jung, Sung-Ju;Park, Mi-Seon;Kim, Jin-Woo;Kim, Yi-Cheong
    • Fisheries and Aquatic Sciences
    • /
    • 제9권4호
    • /
    • pp.146-152
    • /
    • 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.

The First Report of a Megalocytivirus Infection in Farmed Starry Flounder, Platichthys stellatus, in Korea

  • Won, Kyoung-Mi;Cho, Mi Young;Park, Myoung Ae;Jee, Bo Young;Myeong, Jeong-In;Kim, Jin Woo
    • Fisheries and Aquatic Sciences
    • /
    • 제16권2호
    • /
    • pp.93-99
    • /
    • 2013
  • In 2009, a systemic megalocytivirus infection associated with high mortality was detected for the first time in cultured starry flounder Platichthys stellatus in Korea. Diseased starry flounder had pale bodies and gill coloring and enlarged spleens. Histopathological examinations revealed basophilic enlarged cells in various organs of diseased starry flounder. Polymerase chain reaction (PCR) was performed on tissue samples using three published primer sets developed for the red sea bream iridovirus. PCR products were detected for all primer sets, except 1-F/1-R, which are registered by the World Organization for Animal Health (OIE). The part of the gene corresponding to the full open reading frame encoding the viral major capsid protein (MCP) was amplified by PCR. PCR products of approximately 1,581 bp were cloned, and the nucleotide sequences were analyzed phylogenetically. The MCP gene of the starry flounder iridovirus, designated SFIV0909, was identical to that of the turbot reddish body iridovirus (AB166788).

First detection of ranavirus in a wild population of Dybowski's brown frog (Rana dybowskii) in South Korea

  • Park, Jaejin;Grajal-Puche, Alejandro;Roh, Nam-Ho;Park, Il-Kook;Ra, Nam-Yong;Park, Daesik
    • Journal of Ecology and Environment
    • /
    • 제45권1호
    • /
    • pp.10-16
    • /
    • 2021
  • Background: Ranavirus is an emerging infectious disease which has been linked to mass mortality events in various amphibian species. In this study, we document the first mass mortality event of an adult population of Dybowski's brown frogs (Rana dybowskii), in 2017, within a mountain valley in South Korea. Results: We confirmed the presence of ranavirus from all collected frogs (n = 22) via PCR and obtained the 500 bp major capsid protein (MCP) sequence from 13 individuals. The identified MCP sequence highly resembled Frog virus 3 (FV3) and was the same haplotype of a previously identified viral sequence collected from Huanren brown frog (R. huanrenensis) tadpoles in South Korea. Human habitat alteration, by recent erosion control works, may be partially responsible for this mass mortality event. Conclusion: We document the first mass mortality event in a wild Korean population of R. dybowskii. We also suggest, to determine if ranavirus infection is a threat to amphibians, government officials and researchers should develop continuous, country-wide, ranavirus monitoring programs of Korean amphibian populations.

Differentiation of Lymphocystis Disease Virus Genotype by Multiplex PCR

  • Kitamura Shin Ichi;Jung Sung-Ju;Oh Myung-Joo
    • Journal of Microbiology
    • /
    • 제44권2호
    • /
    • pp.248-253
    • /
    • 2006
  • Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. The viruses have been divided into three genotypes (genotype I for LCDV-1, II for Japanese flounder isolates, and III for rockfish isolates) on the basis of major capsid protein (MCP) gene sequences. In this study, we developed a multiplex PCR primer set in order to distinguish these genotypes. We also analyzed the MCP gene of a new LCDV isolate from the sea bass (SB98Yosu). Comparison of sequence identities between SB98Yosu and eight Japanese flounder isolates, revealed identity of more than 90.1 % at nucleotide level and 96.5% at deduced amino acid level, respectively. Phylogenetic analyses based on the MCP gene showed that SB98Yosu belongs to genotype II, along with Japanese flounder isolates. Multiplex PCR based on the MCP gene allowed us to identify these genotypes in a simple and rapid manner, even in a sample that contained two genotypes, in this case genotypes II and III.