• 제목/요약/키워드: major capsid protein

검색결과 40건 처리시간 0.026초

박테리오파지 E3의 Major Capsid Protein을 만드는 유전자의 Mapping 및 염기서열 분석 (Genetic Mapping and Sequence Analysis of the Gene Encoding the Major Capsid Protein of Bacteriophage E3)

  • 배수진;명희준
    • 미생물학회지
    • /
    • 제35권4호
    • /
    • pp.266-269
    • /
    • 1999
  • 박테리오파지 E3가 만드는 plaque은 그 지름이 약 1㎝정도이고 대단히 빠르게 성장한다. 구조 단백질 중 가장 많은 copy를 가지는 major capsid 단백질을 발현하는 유전자를 조절하는 promoter가 가장 효율적일 것이라 생각되며, 이 promoter를 찾기 위하여 먼저 이 유전자를 mapping하였다. 정제한 파지 입자로부터 major capsid 단백질을 분리하여 그 N-terminal amino acid 서열을 확인하였고, 그에 해당하는 degenerate oligonucleotide probe를 이용하여 E3의 genomic library로부터 major capsid 단백질을 발현하는 유전자를 함유하는 clone을 찾았다. 이 clone의 DNA 서열 분석을 통하여 major capsid 단백질을 발현하는 유전자를 확인하였으며, 이는 E3 genome에서 약 72%에 mapping 되었다. 이 gene을 조절하는 promoter의 성질을 고찰하기 위하여 E3의 성장이 rifampicin에 의하여 영향을 받는지 확인한 결과 E3는 자기 고유의 RNA polymerase를 가지고 있음을 알 수 있었다.

  • PDF

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

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.

가잠의 연화병 바이러스에 관한 연구 II. 연화병 바이러스 Polypeptide의 성상 (Studies on Flacherie and Ina-flacherie Viruses of the Silkworm, Bambyx mori II. Some Properties of Polypeptide of Flacherie Virus)

  • 강석권;김근영
    • 한국잠사곤충학회지
    • /
    • 제21권2호
    • /
    • pp.7-10
    • /
    • 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.

  • 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.

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

Negative stain을 이용한 Rotavirus의 투과전자현미경적 구조 (An Electron Microscopic Structure of Rotavirus by Negative Stain)

  • 권중균
    • Applied Microscopy
    • /
    • 제24권4호
    • /
    • pp.107-114
    • /
    • 1994
  • The Rotaviruses are members of the family Reoviridae and are the major cause of severe childhood gastroenteritis worldwide. Recently, electron microscopy has been used to detect non-group A rotaviruses to determine a relatively high resolution structure of the rotavirion. Mature, infectious virions(double-shelled particles) have a diameter of approximately 70nm, and have a capsid structure composed of two concentric protein layers. We have studied patient's stool specimen by negative staining technique complete removal of sucrose suspension. This negative staining technique that could be carried out in about 30 minutes and that could be used with crude stool specimen was an advantage of major significance. Removal of sucrose in the sample by has been completed washing with distilled of sucrose and by washing with distilled water. Ultrastructurally, typical feature of rotavirus has a double capsid construction with an inner capsid of 55nm and on outer 65-70nm diameter can be clearly demonstrated.

  • 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.

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.