• 미생물학회지
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• 제35권4호
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• pp.266-269
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• 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.

• 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.

• 대한바이러스학회지
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• 제26권2호
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• pp.151-162
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• 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.

• 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.

• Journal of Ecology and Environment
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• 제46권4호
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• pp.276-281
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• 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.

• Fisheries and Aquatic Sciences
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• 제16권2호
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• pp.93-99
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• 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).

• 한국어병학회지
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• 제21권3호
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• pp.175-180
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• 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.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2004년도 수산관련학회 공동학술대회 발표요지집
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• pp.413-414
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• 2004

• 생명과학회지
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• 제28권4호
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• pp.478-482
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• 2018

인간 rotavirus는 영아에게 급성 설사를 일으키는 병원체의 하나이다. 본 연구에서는 Escherichia coli의 코돈 선호도를 따라서 인간 rotavirus A (serotype 1 strain WA)의 $VP8^*$ 단백질을 일부분 암호화하도록 인공적인 유전자를 합성하였다. 합성된 $VP8^*$ 유전자는 코돈을 번역틀에 일치시키고 클로닝이 용이하도록 하기 위한 NdeI 및 HindIII 제한효소 절단 부위와 친화적 정제를 위한 6-히스티딘 암호화 서열을 C-말단에 보유하고 있다. 합성된 $VP8^*$ DNA 절편을 pT7-7 발현 벡터에 삽입하여 E. coli BL21 (DE3)로 형질전환한 후에 최종 농도 0.05 mM IPTG로 생산을 유도한 결과 예상했던 대로 19.7-kDa 크기의 $VP8^*$ 단백질이 고농도로 발현되었다. SDS-PAGE에 전개된 단백질들을 대상으로 mouse anti-rotavirus capsid antibody를 사용한 Western blotting의 결과 ~20-kDa $VP8^*$ 단백질 밴드가 관찰되었다. 인공 $Vp8^*$ 단백질이 피하 주사된 토끼의 polyclonal antibody 혈장을 이용한 조사에서도 동일한 크기의 단백질 밴드를 확인할 수 있었다. 이는 합성된 유전자가 바이러스성 질환을 통제할 항원성 백신 후보의 생산 혹은 진단용 항체를 개발하기 위한 쉽고 빠른 방법을 제공할 수 있다는 의미이다.

• Journal of Microbiology
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• 제44권2호
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• pp.248-253
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• 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.