• Title/Summary/Keyword: Vp-p

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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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Effects of Molecular Weight of Poly(4-vinylpyridine) on the Order-Disorder Transition of Molecular Bottle-brush Composed of Poly(4-vinylpyridine) and 3-Pentadecylphenol (P4VP과 PDP로 이루어진 Molecular Bottle-brush의 Order-Disorder Transition에 미치는 P4VP 분자량의 영향에 관한 연구)

  • 최종렬;조항규;전현애;노시태
    • Polymer(Korea)
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    • v.24 no.4
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    • pp.488-498
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    • 2000
  • Molecular bottle-brush was prepared by hydrogen-bonding between poly(4-vinylpyridine)(P4VP) as main chain and 3-pentadecylphenol (PDP) as amphiphilic side chain. Variation of long period ( $L_{p}$), order-disorder transition temperature ( $T_{ODT}$) and mesomorphic structure of bottle-brush were investigated by changing various mole ratio (x) of pyridine group in P4VP and PDP and molecular weight of P4VP. Upper critical solution temperature (UCST) behaviour was observed. For x 0.8-0.9, maximum critical temperature was found. As molecular weight of P4VP was increased, phase transition occurred at higher temperature. It was found that phase behaviour of the bottle-brush was affected by mobility of P4VP as well as size and regularity of lamellar structure. The $L_{p}$ determined from analysis of crystal structure was in the range of 35 $\AA$ and 40 $\AA$ and was more affected by the molecular weight of P4VP than by mole ratio (x). However, if the molecular weight of P4VP was high, LP value was little affected.ted.d.

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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.

Application of AuNPs immobilized on UV Cross-linked P4VP Thin Film as pH Nanosensors (pH 나노센서로의 응용을 위한 UV-가교 P4VP 박막에 고정한 금 나노입자의 특성)

  • Kim, Min-Sung;Jeong, Yeon-Tae
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.21 no.11
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    • pp.1010-1018
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    • 2008
  • In this report, we describe the use of gold nanoparticles (AuNPs) immobilized on pH. responsive, cross-linked poly(4-vinylpyridine) (P4VP) thin films, as a potential application for pH nanosensors. The methodology is based on the variation in surface plasmon resonance of immobilized AuNPs with changing the interparticle distances, caused by the swelling/deswelling of the pH responsive P4VP polymer films. The change in optical properties of the immobilized AuNPs in response to the pH of surrounding media was investigated by a simple yet powerful tool; UV-vis absorption spectroscopy. The swelling of the P4VP chains at pH 2 causes an increase in the interparticle distances of immobilized AUNPS ($\sim20nm$) and hence leads to a blue shift of 48 nm in their surface plasmon resonance band peak. On the other hand, when the surrounding media was altered from pH 2 to 10, a red shift of absorption maxima was observed. The changes were rapid, and the effect was reversible. This system could prove to be useful in fabricating nanosensors for detecting the pH or pH changes of surrounding aqueous medium.

Construction of FMDV VP1 Gene Using Artificial DNA Synthesis and Transformation of Nicotiana tabacum Using Agrobacterium Vector System (유전자 인공합성을 이용한 구제역 유전자 VP1의 제작과 Agrobacterium Vector System을 이용한 담배 형질전환)

  • Lee, Eun-Jung;Lim, Hee-Young;Kim, Sung-Hoon;Kang, Kyung-Sun;Park, Young-Doo;Yun, Choong-Hyo;Yoon, Byoung-Su
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.285-293
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    • 2004
  • FMDV is a viral pathogen that caused foot-and-mouth disease in animals. VP1 is a major capsid protein of FMDV. It is known as one of best materials for the FMDV diagnosis and for the development of protein vaccine. In this study, 633 bp of VP1 gene was modified for the expression of VP1 in plant, based on the VP1 DNA sequence from FMDV taiwan O type and from FMDV isolated vietnam. The. deduced DNA fragment was artificially synthesized using the multiple fragment extension with long-nucleotides. A new plant transgenic vector system, pCAMBIA139011 was constructed on the basis of pBI12l and pCAMBIA1390. Using this vector system and GFP gene or modified VP1 gene, each target gene was introduced into Nicotiana tabacum. The insertion of whole target gene was successfully confirmed in each transgenic plant named GFP-A7 and VP1-4, respectively. The expression level of each gene was estimated by RT-PCR and Real-Time PCR using VP1, GFP specific primers.

Investigation of Tube Voltage Range using Dose Comparison based on Effective Detector Exposure Index in Chest Radiography (흉부 X-ray 검사 시 선량 비교를 활용한 유효 Detector Exposure Index 기반의 적절한 관전압 범위 제안)

  • Shim, Jina;Lee, Youngjin
    • Journal of the Korean Society of Radiology
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    • v.15 no.2
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    • pp.139-145
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    • 2021
  • This study is to confirm the range of tube voltage for Chest X-ray in DR system by comparing with dose area product (DAP) and effective dose in efficient detector exposure index (DEI) range. GE definium 8000 was used to for the phantom study. The range of tube voltage is 60~130 kVp and of mAs is 2.5~40 mAs. The acquired images were classified into efficient DEI groups, then calculated effective dose with DAP by using a PC-Based Monte Carlo Program 2.0. The signal to noise ratio (SNR) was measured at 4 regions, including the thoracic spine, the lung area with the ribs, the lung area without the ribs, and the liver by using Picture Archiving and Communication System. The significance of the group for each tube voltage was verified by performing the kruskal-wallis test and the mann-whitney test as a post-test. When set to 4 groups dependned on the tube voltage, DAP showed significant differences; 60 kVp and 80 kVp, and 60 kVp and 90 kVp (p= 0.034, 0.021). Effective dose exhibited no statistically significant differences from the all of the group (p>0.05). SNR exhibited statistically significant differences from the all of the group in the liver except compared to 80 kVp and 90 kVp (p<0.05). Therefore, high tube voltages of 100 kVp or more need to be reconsidered in terms of patient dose and imaging in order to represent an appropriate chest X-ray image in a digital system.

Fabrication of an Alternating Multilayer Film of Poly(ethylene-alt-maleic anhydride) and Poly(4-vinyl pyridine) by Layer-by-Layer Self-Assembly Method (Layer-by-Layer 자기조립법에 의한 Poly(ethyiene-alt-maleic anhydride)i Poly(4-vinyl pyrtdine) 다층막 제조)

  • Lee Joon Youl;Hong Sook-Young
    • Polymer(Korea)
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    • v.29 no.4
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    • pp.392-398
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    • 2005
  • Self-assembled multilayer thin films of poly(ethylene-alt-maleic anhydride) (PEMAh) and poly(4-vinyl pyridine) (P4VP) were fabricated by layer-by-layer (LbL) sequential adsorption. Fourier transform infrared (FT-IR) spectroscopic analysis of the self-assembled PEMAh/P4VP multilayer films confirms that the driving forces for the multilayer buildup are the intermolecular hydrogen bonding and electrostatic interactions. The linear increase of absorption peak of P4VP at 256 nm with increasing number of PEMAh/P4VP bilayers indicates that the multilayer buildup is an uniform assembling process. We also investigate the effects of polyelectrolyte concenhation variation of the dipping solution and pH variation of the PEMAh solution on the multilayer film formation. Thickness. adsorbed polyelectrolyte mass and surface roughness of the multilayer films were measured by UV-visible spectroscopy, quartz crystal microbalance (QCM), and atomic force microscopy (AFM), respectively.

Effect of PEO of PS-P2VP photonic gel films

  • Shin, Sung-Eui;Kim, Su-Young;Shin, Dong-Myung
    • 한국정보디스플레이학회:학술대회논문집
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    • 2009.10a
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    • pp.1405-1407
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    • 2009
  • We prepared polystyrene-b-poly(2-vinyl pyridine) (PSb-P2VP) lamellar films which is hydrophobic block-hydrophilic polyelectrolyte block polymer have 57 kg/mol-b-57 kg/mol. The result of UV-visible absorption spectra supported that effect of poly(ethylene oxide) on the band gap tuning of PS-P2VP photonic gel like salt effect.

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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.

Studies on the VP4 and VP7 Genes of Bovine Rotaviruses from Field Samples Using RT-PCR and RFLP Analysis (RT-PCR과 RFLP법을 이용한 국내 소 로타바이러스 VP4 및 VP7 유전자의 특성 규명)

  • Jeon, Seong-Jin;Chang, Chung-Ho;Chung, Chung-Won;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.2
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    • pp.165-174
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    • 1998
  • Characterizations of the VP4 (P type) and VP7 (G type) genes of Korean isolates of bovine rotavirus were performed using RT-PCR/RFLP and nucleotide sequencing analysis. After RT-PCR amplification of partial length (1094bp) of the VP4 and full length (1062bp) of the VP7 genes, amplified PCR products were digested with restriction endonucleases and digestion patterns were compared with those of reference rotaviruses. With the VP4 genes, four RFLP (A-D) profiles were observed; three (A, Band C) were the same as those of bovine rotavirus NCDV (P[1]), IND (P[5]) and B223 (P[11]), respectively. Profile D was the same as that of porcine rotavirus OSU (P[7]). With the VP7 genes, five RFLP profiles (I-V) were observed; three of them (I, II and III) were the same as those of bovine rotavirus NCDV (G6), Cody 1-801 (G8), and B223 (G10), respectively. Profile IV and V were atypical to those of reference bovine rotaviruses used in this study. These two profiles were identified as G6 and G5, respectively, after analyzing and comparing the nucleotide sequences. The G typing analysis revealed that 61.9% (26/42) were G6, which included G6 subtype; 28.6% (12/42) were G5; 7.1% (3/42) were G10; 2.4% (1/42) were G8. The P typing analysis revealed that 54.8% (23/42) were P[5]; 28.6% (12/42) were P[7]; 11.8% (5/42) were [11]; 4.8% (2/42) were P[1]. Our results showed that G6/P[5] were the most prevalent rotaviruses in diarrheic calves in Korea. Also, this is the first report that G5/P[7] rotaviruses were identified from cattle with diarrhea.

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