• Title/Summary/Keyword: VP6

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Expression of Rotavirus Capsid Proteins VP6 and VP7 in Mammalian Cells Using Semliki Forest Virus-Based Expression System

  • Choi, Eun-Ah;Kim, Eun;Oh, Yoon-I;Shin, Kwang-Soon;Kim, Hyun-Soo;Kim, Chul-Joong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.463-469
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    • 2002
  • Rotaviruses are the world-wide leading causative agents of severe dehydrating gastroenteritis in young children and animals. The outer capsid glycoprotein VP7 and inner capsid glycoprotein VP6 of rotaviruses are highly antigenic and immunogenic. An SFV-based expression system has recently emerged as a useful tool for heterologous protein production in mammalian cells, exhibiting a much more efficient performance compared to other gene expression systems. Accordingly, the current study adopted an SFV-based expression system to express the VP7 of a group A human rotavirus from a Korean isolate, and the VP6 of a group B bovine rotavirus from a Korean isolate, in mammalian cells. The genes of the VP6 and VP7 were inserted into the SFV expression vector pSFV-1. The RNA was transcribed in vitro from pSFV-VP6 and pSFV-VP7 using SP6 polymerase. Each RNA was then electroporated into BHK-21 cells along with pSFV-helper RNA containing the structural protein gene without the packaging signal. The expression of VP6 and VP7 in the cytoplasm was then detected by immunocytochemistry. The recombinant virus was harvested by ultracentrifugation and examined under electron microscopy. After infecting BHK-21 cells with the defective viruses, the expressed proteins were separated by SDS-PAGE and analyzed by a Western blot. The results indicate that an SFV-based expression system fur the VP6 and VP7 of rotaviruses is an efficient tool for developing a diagnostic kit and/or preventive vaccine.

Molecular characterization of avian rotavirus isolated in Korea

  • Wang, Jun-Hui;Koo, Bon-Sang;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.1
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    • pp.23-30
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    • 2013
  • An avian rotavirus (AvRV-2) was isolated from feces of broilers suffering from acute gastroenteritis in 2011. It was the first avian rotavirus isolated in Korea. To investigate the molecular characteristics of AvRV-2, the VP4, VP6, VP7 and NSP4 gene nucleotide sequences were determined and compared with those of rotavirus strains available in the GenBank database. The phylogenetic tree of VP7 gene showed that AvRV-2 had a high degree of nucleotide sequence homology (93.4% to 94.7%) with those of rotaviruses belonging to genotype G19 cluster. The phylogenetic tree of the VP4 gene revealed a high degree of nucleotide sequence homology (95.8% to 95.9%) with genotype P[30] rotaviruses isolated from chickens. The VP6 and NSP4 gene nucleotide sequences showed the highest identities with those of avian strains with 95.3% to 96.4% and 90.3% to 92.2%, respectively. Genetic characterization of the VP4, VP6, VP7 and NSP4 showed that AvRV-2 strain was most closely related to chicken rotavirus strains from Germany and Japan. Comparative nucleotide sequences and phylogenetic analysis indicated that avian rotavirus isolated from broilers belonged to genotype G19P[30] and it was the first report on avian rotavirus infection in Korea.

Recombination and Expression of VP1 Gene of Infectious Pancreatic Necrosis Virus DRT Strain in a Baculovirus, Hyphantria cunea Nuclear Polyhedrosis Virus (전염성 췌장괴저바이러스 DRT Strain VP1유전자의 Baculovirus Hyphantria cunea Nuclear Polyhedrosis Virus에 재조합과 발현)

  • Lee, Hyung-Hoan;Chang, Jae-Hyeok;Chung, Hye-Kyung;Cha, Sung-Chul
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.239-255
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    • 1997
  • Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

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Production and diagnostic application of monoclonal antibodies against infectious bursal disease virus (IBDV에 대한 단크론항체 생산 및 진단적 응용)

  • Ryu, Min-Sang;Song, Yoon-Ki;Lee, Seung-Chul;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.5-12
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    • 2011
  • Infectious bursal disease (IBD) caused by infectious bursal disease virus (IBDV) is a highly contagious viral disease in chicken. It causes heavy economic loss by immune suppression and high mortality. The IBDV, designated Avibirnavirus in the Family Birnaviridae, has a double-stranded RNA genome formed by two segments, segment A and segment B. Segment A encodes a 108 KDa polypeptide that is self-cleaved to produce pVP2, VP3 and VP4, and later pVP2 is cleaved to VP2. The VP2 contains the antigenic regions responsible for elicitation of neutralizing antibodies and VP3 is a major immunogenic protein of IBDV. In this study, monoclonal antibodies (MAbs) specific for IBDV were produced and characterized. All 15 MAbs were specific for IBDV and did not react with other viruses used in this study. The protein specificity of MAbs was determined by comparing the reactivity patterns of each MAb with IBDV VP2 and VP234 recombinant baculoviruses and Western blot analysis. As a result, 7 MAbs (1F5, 2C8, 2F4, 3C7, 4C3, 6F11, 6G5) and 5 MAbs (2A4, 2G2, 3F5, 3G2, 4F10) were specific for VP2 and VP3, respectively. The protein specificity of 3 MAbs (2B8, 3F7, 3F8) were not determined. Five (2C8, 2F4, 4C3, 6F11, 6G5) of the VP2-specific MAbs had a neutralizing activity against IBDV. Some MAbs reacted with IBDV-infected bursa of Fabricius by indirect fluorescence antibody (IFA) and immunohistochemistry (IHC) assay. The MAbs produced in this study would be used for diagnostic reagents for the detection of IBDV infection.

Characteristics and Influence of Scattering Radiation in Cultural Heritage Radiography (문화재 방사선 조사에서 발생하는 산란 방사선의 특성과 영향)

  • Song, Jung Il;Park, Young Hwan;Yu, Ji Hye
    • Journal of Conservation Science
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    • v.34 no.6
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    • pp.539-548
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    • 2018
  • This study was conducted to evaluate the effects of scattering radiation, which was not considered in the cultural Heritage radiography, by evaluating the relationship between the tube voltage (unit: kVp), film-floor-distance(FFD), and lead screen layout. The density (unit: D) of the test specimens and the scattered radiation increased with the tube voltage. The density of the test specimens showed an average deviation of 1.4 D; it was 0.17 D at 60 kVp, 1.54 D at 160 kVp, and 2.97 D at 220 kVp. The mean density of the scattered radiation was 0.10 D at 60 kVp, 0.40 D at 160 kVp, and 0.46 D at 220 kVp. The density tended to increase when the tube voltage ranged between 60 kVp and 160 kVp, as the FFD distance increased. However, a change in the permeation density was not observed for high voltages(160 kVp-220 kVp). Scattered radiation was observed when FFD was 50 mm, 100 mm, and 200 mm and no lead screen was used and the bottom surface was replaced with the lead screen. No scattered radiation was observed when FFD was 0 mm. The identification rate ranged from 2.08% to 2.67%, according to the FFD, for a 160 kVp tube voltage, and from 2.67% to 3.33% for a 220 kVp tube voltage.

Vp28 of Shrimp White Spot Syndrome Virus Is Involved in the Attachment and Penetration into Shrimp Cells

  • Yi, Guohua;Wang, Zhimin;Qi, Yipeng;Yao, Lunguang;Qian, Juan;Hu, Longbo
    • BMB Reports
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    • v.37 no.6
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    • pp.726-734
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    • 2004
  • White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

Coding Efficiency Comparison between Next Generation Video Codecs: HEVC vs VP9 (차세대 동영상 코덱 압축 효율 비교: HEVC vs VP9)

  • Kim, Il-Koo
    • Proceedings of the Korean Society of Broadcast Engineers Conference
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    • 2013.06a
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    • pp.176-179
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    • 2013
  • 본 논문에서는 JCT-VC 에서 2013 년 1 월에 표준화가 완료된 High Efficiency Video Coding (HEVC)과 구글에서 2013 년 6 월에 개발 완료 예정인 VP9 의 압축 효율 비교를 수행한다. HEVC 는 UHD 등 고화질 방송 등에 대응하도록 디자인 되었으며, VP9 은 유튜브 (YouTube) 등과 같은 인터넷 비디오 스트리밍에 적합하도록 디자인되었다. VP9 의 경우 HEVC 와는 달리 로열티 프리 (royalty-free)를 지향하며 오픈소스 (open source) 방식으로 개발이 진행되고 있다. 본 논문에서는 HEVC 와 VP9 의 디자인 차별점을 소개하고, 랜덤 액세스 환경(Random Access, RA)과 저지연 환경 (Low Delay, LD)에서 HEVC 와 VP9 의 압축 효율을 비교한다. 실험 결과에 따르면, 방송 및 패키지 미디어 등에서 많이 사용될 랜덤 액세스 환경에서는 VP9 이 HEVC 대비 32.7% 열세를 보인다. 비디오 컨퍼런스등과 같은 저지연 환경에서는 VP9 이 HEVC 대비 26.7% 열세를 보인다. VP9 의 경우 개발이 완료된 것이 아니므로, 향후 압축 효율의 향상이 있을 것으로 기대된다.

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Oxidation of Ascorbic Acid by Crosslinked Poly(4-vinyl pyridine)-Cu(II) Complexes 2. Effect of Crosslinker (가교 폴리(4-비닐피리딘)-구리(II) 착물에 의한 Ascorbic Acid의 산화반응 2. 가교제의 영향)

  • 이석기;서재곤;구광모;전일련;김우식
    • Polymer(Korea)
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    • v.24 no.2
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    • pp.252-258
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    • 2000
  • Various crosslinked poly(4-vinylpyridines) (CHP4VP) having different degrees of crosslinking were synthesized by radical copolymerization of 4-vinylpyridine with if N,N' -1, 6-hexamethylenebisacrylamide, and CHP4VP- Cu(II) complexes were prepared by the method of adsorption equilibrium. The catalytic activity of the complexes for the oxidation of ascorbic acid (AA) was investigated. The oxidation of AA by these complexes showed a kinetic behavior of the Michaelis-Menten type. The catalytic activity of CHP4VP-Cu(I ) catalytic system was increased with increasing the degree of crosslinking of CHP4VP, and its activity was scarcely decreased even after repeated use. However, the tendency of the catalytic activity of CHP4VP-Cu(II) complexes was decreased for the oxidation of AA when compared with that of the previously reported catalytic system containing crosslinked poly(4-vinylpyridine) prepared using N,N'-methylenebisacrylamide as a crosslinker. These results indicate that the degree of crosslinking of CHP4VP and the hydrophobicity of the crosslinker play an important role in the catalytic system of the oxidation of AA.

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Design of Unified HEVC/VP9 4×4 Transform Block (HEVC/VP9 4×4 Transform 통합 블록 설계)

  • Jung, Seulkee;Lee, Seongsoo
    • Journal of IKEEE
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    • v.19 no.3
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    • pp.392-399
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    • 2015
  • This paper proposes a unified $4{\times}4$ transform architecture for HEVC and VP9 codec to reduce hardware size. It performs HEVC $4{\times}4$ IDCT, HEVC $4{\times}4$ IDST, VP9 $4{\times}4$ IDCT, and VP9 $4{\times}4$ IADST in a unified hardware. HEVC $4{\times}4$ IDCT and VP9 $4{\times}4$ IDCT have same IDCT computation except for the scales of coefficients. Similarly, HEVC $4{\times}4$ IDST and VP9 $4{\times}4$ IADST have same IDST computation except for the scales of coefficients. Furthermore, IDCT and IDST have quite a lot of similarity, so they can share some hardwares in common. So the proposed hardware performs all 4 operations in a unified hardware, where each operation has its own multiplication coefficients with shared butterfly adders. The synthesized block in 0.18 um technology is 6,679 gates, and the gate count is reduced by 25.3% in comparison with conventional designs.

Genetic characterization of bovine rotavirus isolates in Korea

  • Yang, Dong-Kun;Kim, Byoung-han;Lee, Kyung-Woo;Kim, Yeon-Hee;Song, Jae-Young;Park, Joong-Won;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.423-429
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    • 2008
  • Throughout the world, rotavirus infections cause extensive morbidity in human infants and diarrhea in animals such as white scour caused by bovine rotavirus in calves. We isolated three rotavirus strains designated KV0407, KV0418, and KV0426 from 103 fecal samples of diarrheic calves. The genes coding for proteins VP4, VP6, VP7, and NSP4 from strain KV0407 were sequenced and compared with the nucleotide sequences of other known strains of rotavirus. The KV0407 VP4 gene was highly homologous to the OSU (99.4%) and JL94 (99.4%), but not the B223 (62.4%) and K33 (62.4%) VP4 genes. The KV0407 and KV0418 VP7 genes were most similar to the OSU and super-short type VMRI VP7 genes. Based on nucleotide sequence analysis, the KV0407 strain was tentatively assigned to A serogroup (SG I), G5P[7], NSP4 genotype B and the KV0418 and KV0426 strains were assigned to A serogroup (SG I), G6P[5], NSP4 genotype A. The genetic characterization of these bovine rotavirus isolates could be useful for the diagnosis and prevention of diarrhea in calves.