• Title/Summary/Keyword: nucleocapsid protein

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Unusual Features of Human Immunodeficiency Virus Type-1 Virion (면역결핍 바이러스 입자의 비특이적 성질)

  • Shin, Cha-Gyun
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.107-114
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    • 1996
  • 본 연구는 인간면역결핍바이러스의 입자를 비이온성 계면활성제로 처리할 때 바이러스 입자구조에서 분리되어 방출되는 바이러스 구조단백질들의 분포를 sucrose gradient로 분석하여, 바이러스 입자를 구성하는 바이러스 구조단백질과 바이러스입자의 생물리학적 특성을 연구하였다. 바이러스입자들을 0.16% NP40 (Nonidet P-40)으로 처리할 때, 바이러스 capsid 단백질과 바이러스 막 단백질 (membrance protein)들은 다른 바이러스 구성성분들과 잘 분리되었다. 계면활성제처리에서 방출되지 않은 구성 성분들은 matrix 단백질, nucleocapsid 단백질, reverse transcriptase, integrase 및 바이러스 RNA genome로써, 이들은 subviral 구조를 형성한다. 이러한 결과는 상대적으로 다른 바이러스들의 capsid 단백질과 면역 결핍 바이러스의 capsid 단백질 (p24)를 비교할 때, 면역결핍바이러스의 capsid 단백질은 바이러스핵을 형성할 때, capsid 단백질 사이의 결합력이 매우 약한 것으로 추정된다. 또한 바이러스 조절단백질의 하나인 vpr 단백질을 함유하는 바이러스입자를 NP40 처리하여 분석하였을 때, vpr 단백질은 subviral 구조에 존재하는 것으로 나타났다.

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Molecular detection of bat coronaviruses in three bat species in Indonesia

  • Dharmayanti, Ni Luh Putu Indi;Nurjanah, Diana;Nuradji, Harimurti;Maryanto, Ibnu;Exploitasia, Indra;Indriani, Risa
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.70.1-70.12
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    • 2021
  • Bats are an important reservoir of several zoonotic diseases. However, the circulation of bat coronaviruses (BatCoV) in live animal markets in Indonesia has not been reported. Genetic characterization of BatCoV was performed by sequencing partial RdRp genes. Real-time polymerase chain reaction based on nucleocapsid protein (N) gene and Enzyme-linked immunosorbent assay against the N protein were conducted to detect the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA and antibody, respectively. We identified the presence of BatCoV on Cynopterus brachyotis, Macroglossus minimus, and Rousettus amplexicaudatus. The results showed that the BatCoV included in this study are from an unclassified coronavirus group. Notably, SARS-CoV-2 viral RNA and antibodies were not detected in the sampled bats.

A Cell-based Method to Monitor the Interaction between Hepatitis B Virus Capsid and Surface Proteins

  • Kim, Yun-Kyoung;Oh, Soo-Jin;Jin, Bong-Suk;Park, Chan-Hoo;Jeon, Hye Sung;Boo, Doo-Wan;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.30 no.3
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    • pp.577-581
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    • 2009
  • Interactions between the surface and capsid proteins of the hepatitis B virus (HBV) are critical for the assembly of virus particles. In this study, we developed a cell-based method to visualize the interactions between the capsid and surface proteins of HBV. Capsid-GFP, a capsid protein fused to a green fluorescence protein (GFP), forms nucleocapsid-like structures in the cytoplasm of mammalian cells. It relocates to the plasma membranes in cells expressing PH-PreS, a fusion protein consisting of the PreS region of the HBV surface protein and the PH domain of PLC-$\gamma$. Membrane localization of the capsid-GFP in these cells is prevented by an inhibitory peptide that blocks the interaction between the capsid and surface proteins. This dynamic localization of capsid-GFP is applicable for screening compounds that may potentially inhibit or prevent the assembly process of HBV particles.

Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • Nam, Ki-Yean;Chung, Dong-Hoon;Choi, Je-Won;Lee, Yun-Seong;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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Clearance of False-positive Antigen-Antibody Reactions of a Diagnostic Antigen Production in Escherichia coli with Human Sera

  • Noh, Kap-Soo;Kim, Jong-Wan;Ha, Suk-Hoon;Yoo, Wang-Don;Jeon, Weong-Joong;Kim, Hyun-Su
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.63-65
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    • 1999
  • Although many pharmaceutically useful proteins are produced in E. coli expression system, it is very are rare for the system to be used in the production of diagnostic antigen due to a major problem, i.e., false-positive reaction of e. coli host-derived proteins contaminating purified diagnostic antigen with human sera. The N (nucleocapsid) protein of Seoul virus causing haemorrhagic fever with renal syndrome (HFRS) was produced in E. coli BL21 (DE3), and used for the detection of N protein-specific antibodies in human sera. Using the N protein as a diagnostic antigen of HFRS, the false-positive reaction was cleared by merely mixing the test sera with the extract of E. coli host strain not harboring expression plasmid.

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Biochemical Characteristics of Spodoptera exigua Nuclear Polyhedrosis Virus (파밤나방 핵다각체병 바이러스의 생화학적 특성)

  • 진병래;박범석;재연호;강석권
    • Korean journal of applied entomology
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    • v.30 no.2
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    • pp.144-149
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    • 1991
  • Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

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Hepatitis C Virus Core Protein Is Efficiently Released into the Culture Medium in Insect Cells

  • Choi, Soo-Ho;Kim, So-Yeon;Park, Kyu-Jin;Kim, Yeon-Joo;Hwang, Soon-Bong
    • BMB Reports
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    • v.37 no.6
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    • pp.735-740
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    • 2004
  • Hepatitis C virus (HCV) is a causal agent of the chronic liver infection. To understand HCV morphogenesis, we studied the assembly of HCV structural proteins in insect cells. We constructed recombinant baculovirus expression vectors consisting of either HCV core alone, core-E1, or core-E1-E2. These structural proteins were expressed in insect cells and were examined to assemble into particles. Neither core-E1 nor core-E1-E2 was capable of assembling into virus-like particles (VLPs). It was surprising that the core protein alone was assembled into core-like particles. These particles were released into the culture medium as early as 2 days after infection. In our system, HCV structural proteins including envelope proteins did not assemble into VLPs. Instead, the core protein itself has the intrinsic capacity to assemble into amorphous core-like particles. Furthermore, released core particles were associated with HCV RNA, indicating that core proteins were assembled into nucleocapsids. These results suggest that HCV may utilize a unique core release mechanism to evade the hosts defense mechanism, thus contributing to the persistence of HCV infection.

p13 from group II baculoviruses is a killing-associated gene

  • Lu, Nan;Du, Enqi;Liu, Yangkun;Qiao, Hong;Yao, Lunguang;Pan, Zishu;Lu, Songya;Qi, Yipeng
    • BMB Reports
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    • v.45 no.12
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    • pp.730-735
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    • 2012
  • p13 gene was first described in Leucania separata multinuclear polyhedrosis virus (Ls-p13) several years ago, but the function of P13 protein has not been experimentally investigated to date. In this article, we indicated that the expression of p13 from Heliothis armigera single nucleocapsid nucleopolyhedrovirus (Ha-p13) was regulated by both early and late promoter. Luciferase assay demonstrated that the activity of Ha-p13 promoter with hr4 enhancer was more than 100 times in heterologous Sf9 cells than that in nature host Hz-AM1 cells. Both Ls-P13 and Ha-P13 are transmembrane proteins. Confocal microscopic analysis showed that both mainly located in the cytoplasm membrane at 48 h. Results of RNA interference indicated that Ha-p13 was a killing-associated gene for host insects H. armigera. The AcMNPV acquired the mentioned killing activity and markedly accelerate the killing rate when expressing Ls-p13. In conclusion, p13 is a killing associated gene in both homologous and heterologous nucleopolyhedrovirus.

Molecular Cloning and Nucleotide Sequence of the N Protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus (한국에서 분리된 전염성 조혈괴저바이러스의 N 단백질의 유전자 클로닝과 염기서열 분석)

  • Mun, Chang-Hoon;Kim, Hyun-Ju;Park, Jeong-Min;Cho, Wha-Ja;Cha, Seung-Ju;Yoon, Won-Joon;Park, Jeong-Jae;Lee, Eun-Hee;Kang, Hoe-Sung;Kim, Han-Do;Park, Jeong-Woo
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.69-73
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    • 1998
  • We have cloned and analyzed cDNA coding for nucleocapsid protein N from infectious hematopoietic necrosis virus(IHNV), IHNV-PRT, which was isolated in Korea. The N gene had open reading frame of 1,176 bp that encoded a 391 amino acids with a molecular weight of 42.3 kDa. The deduced amino acid sequence of N protein was 75-90% identical to those of foreign isolates, IHNV, but was 43% and 38% identical to those of other species of fish rhabdovirus, hirame rhabdovirus(HRV) and viral hemorrahagic septicemia virus(VHSV), respectively. However, it revealed high levels of sequence identity between 214-265 amino acid sequences among all species of fish rhabdovirus.

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Occurrence of Tomato spotted wilt virus in Chrysanthemum (Dendranthema grandiflorum) in Korea

  • Chung Bong-Nam;Pak Ha-Seung;Jung Jae-A;Kim Jeong-Soo
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.230-234
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    • 2006
  • Tomato spotted wilt virus (TSWV) has been identified in commercial chrysanthemum cultivars in Korea. Nucleotide sequences of the N gene of TSWV-ch14 isolated from infected chrysanthemum were determined and deposited in GenBank under accession no. DQ453158. The symptoms consisted of dark colored leaf necrosis, black streaks along the stem, wilting of plant parts in 'Sinma'; and chlorotic spots, necrosis of axillary shoots and withering of leaves in 'Hwarang'. Electron micrographs of leaf preparation of Nicotiana rustica infected with TSWV-ch14 contained spherical particles around 85 nm in diameter. TSWV was identified from chrysanthemum by sequence determination of N nucleocapsid protein and virion observation by transmission electron microscope. This is the first reported observation on TSWV in chrysanthemum in Korea.