• Title/Summary/Keyword: VP2

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Expression of Recombinant Rotavirus Proteins Harboring Antigenic Epitopes of the Hepatitis A Virus Polyprotein in Insect Cells

  • Than, Van Thai;Baek, In-Hyuk;Lee, Hee-Young;Kim, Jong-Bum;Shon, Dong-Hwa;Chung, In-Sik;Kim, Won-Yong
    • Biomolecules & Therapeutics
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    • v.20 no.3
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    • pp.320-325
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    • 2012
  • Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.

Characterization of G9 genotype porcine rotavirus isolated in Korea (국내 양돈장에서 분리한 G9형 돼지로타바이러스의 특성)

  • Wang, Jun-Hui;Lee, Seung-Chul;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.83-90
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    • 2012
  • Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.

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.

Production of monoclonal antibodies against VP28 of white spot syndrome virus (WSSV) (White spot syndrome virus (WSSV)의 VP28에 대한 단클론 항체 생산)

  • Bang, Ji-hyeong;Kim, Wi-Sik;Kim, Choon-sup;Kim, Jong-Oh;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.32 no.1
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    • pp.45-48
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    • 2019
  • We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.

A Study on the Preparation of Carboxylated Polysulfone/MeVpI-DVB Membranes and Its Characteristics (Carboxylated Polysulfon/MeVpl-DVB 막의 제조와 특성에 관한 연구)

  • 김관식;전경용;조영일
    • Membrane Journal
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    • v.5 no.1
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    • pp.26-34
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    • 1995
  • In this study, polysulfone was carboxylated(CPSf), as a method of introducing carboxyl group to polymer main chain using direct lithiation reaction. Then, poly(1-alkyl-4-vinylpyridinium iodide-co-divinylbenzene) (MeVpI-DVB) containing pyridinium cation which has an anion selectivity as a fixed carrier was synthesized. And polymer membranes were prepared by mixing CPSf and MeVpI-DVB. Characteristics and permeation of membranes were investigated. As a result of synthesizing CPSf/MeVpI-DVB, blend was formed, not new copolymer. As the content of CPSf amount increasing, thermal stability of membranes was increasing. Ion exchange capacity was 1.0~1.8(meq/g dry mem.) and water content was 0.16~0.26(g $H_2{O}$)/g dry mem.) and fixed ion concentration was 6.4~7.3(meq/g $H_2{O}$) in synthetic membranes. The $Cl^-$ flux showed an increase due to the increase of CPSf content.

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Effects of Vacuum Precooling on Shelf Life of Pleurotus eryngii during PE Packaging Storage (큰느타리 버섯의 PE 포장 저장 중 선도에 미치는 예냉처리 효과)

  • Beik, Kyung-Yean;Lee, Ye-Kyung;Kim, Jae-Won;Park, In-Sik;Kim, Soon-Dong
    • Food Science and Preservation
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    • v.16 no.2
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    • pp.166-171
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    • 2009
  • The effects of vacuum precooling(VP) on the shelf-life of polyethylene film(PE) packaged King oyster mushrooms(Pleurotus eryngii) during storage at $-1^{\circ}C$ were investigated. VP was conducted below $0^{\circ}C$ in a $-1^{\circ}C$ cold chamber of 40 minutes, and mushrooms were stored for 30 days in batches of 1kg. The weight loss of the VP-treated mushroom was slightly lower than that of control. The $O_2$ concentrations of VP-treated mushroos, within 4 days of storage, were 2.44-14.50 %/kg-package/hr, thus higher than control values(2.01-8.19%/kg-package/hr). $CO_2$ generation of VP-treated mushrooms, again within 4 days of storage, was 0.47%/kg-package/hr, thus lower than that of controls(0.58%/kg-package/hr). The $CO_2/O_2$ ratio peaked on day 4 of storage in the control group, tbut no such peak was observed in VP-treated mushrooms. In the VP-treated fungi, lightness was higher, and redness and yellowness lower, than in controls, at all storage times.. In VP-treated mushrooms, strength, hardness and chewiness were significantly higher than in controls, but there were no significant differemces in springiness or cohesiveness. Softening and breakdown of under-cap wrinkles were observed in control mushrooms stored for 30 days, but occurred to a lesser extent in VP-treated fungi. Stipe reticulum tissue vessels in the 30 day-stored VP-treated mushrooms were relatively well-defined and clear, but were softer and diffuse in the control fungi. The results thus confirmed that VP after harvest enhanced mushroom shelf-life and PE packaging prolonged storage time. The data will have industrial applications.

Resistance against white spot syndrome virus (WSSV) infection in wild marine crab Gaetice depressus by injection of recombinant VP28 protein

  • Kim, Chun Soo;Choi, Seung Hyuk;Kim, Min Sun;Kim, Ki Hong
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.11-16
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    • 2014
  • The resistance against white spot syndrome virus (WSSV) infection in wild marine crab Gaetice depressus by the immunization of a recombinant glutathione-S-transferase (GST) fused VP28 protein (GST-VP28) was evaluated. The cumulative mortalities of GST-VP28 injected groups were lower than those of the control groups at 10 days of post-challenge, and the time to death of 50% crab ($TD_{50}$) was delayed by the immunization using GST-VP28. The group boosted with GST-VP28 after 2 weeks of primary immunization clearly showed longer $TD_{50}$ than non-boosted group against challenge with WSSV. This result suggests that boosting with the antigen protein elicit stronger immune responses similar to adaptive immune responses of vertebrates. However, the short $TD_{50}$ was observed in the group challenged at 3 weeks post boosting comparing to the group challenged at 1 week post boosting. This suggests that the protective strength of immunization decreased by the time.

Quality Changes of Pork in Relation to Packaging Conditions During Chilled Storage in Households

  • Lee, Keun Taik;Jang, Min Jun
    • Food Science of Animal Resources
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    • v.33 no.4
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    • pp.448-455
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    • 2013
  • Proper storage of left-over meat in a household refrigerator is important for extending its shelf-life and assuring its safety until it is next used. Various fresh meat packaging methods were examined to determine their effects on the quality characteristics of pork loins during household storage at $5^{\circ}C$. The packaging methods include 1) wrapping in a polyethylene pouch (WP), 2) keeping in an air-tight plastic container (CP), and 3) using a household vacuum packaging machine (VP). The fastest increase in total aerobic bacteria during storage occurred in the WP samples, followed by the CP and VP samples. The count of Pseudomonas spp. was found to be lowest in the VP, and then the CP and WP samples. Enterobacteriaceae grew fastest in the WP samples, followed by the CP and VP samples. The WP samples also incurred the most significant increase in thiobarbituric acids and volatile basic nitrogen values over the storage period, as compared to the CP and VP samples. Off-odour at 30 min after opening the package, was first notable at day 11 in WP samples, but only at day 15 in the CP and VP samples. The colour also deteriorated earlier in the WP samples than in the CP and VP samples.

Typing and Sequence Analysis of the VP7 Gene of Rotavirus Isolated from Infants in Korea (한국영아에서 분리된 로타바이러스의 VP7 유전자형 및 염기서열 분석)

  • Song, Mi-Ok;Yun, Karyn;Chung, Sang-In;Choi, Chul-Soon;Lim, In-Seok;Kang, Shien-Young;An, Chang-Nam;Kim, Won-Yong
    • The Journal of Korean Society of Virology
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    • v.30 no.2
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    • pp.101-112
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    • 2000
  • Rotaviruses are the most common cause of severe vomiting and diarrhea in children worldwide and classified as a genus in the family Reoviridae. Rotavirus has eleven segmented dsRNAs and the virion consists of three shells. Outer capsid VP7 and VP4 induce neutralizing antibodies and are classified into G types (glycoprotein VP7) and P types (protease-sensitive VP4). Characterization of VP7 gene of Korean isolates of human rotavirus was performed using multiplex PCR and nucleotide sequence analysis. After RT-PCR amplification of full length (1,062 bp) of VP7 genes, the amplified PCR products were G typed by multiplex PCR and the nucleotide sequences were compared with those of reference rotavirus from GenBank. The G type analysis revealed that 25% (2/8) belong to G1, whereas 37.5% (3/8) benong to G2 and G4, respectively. The Korean isolates within the same serotypes showed high homology of nucleotide sequences and could be discriminated from foreign isolates exception with two strains (CAU009 and CAU022). But Korean isolates CAU009 and CAU022 were close related into japanease isolates 417 (99.2%) and indian isolates (97.6%) than Korean isolatese. Our results showed that these two strains were supposed to be originated from abroad. As a results, The G typing and nucleotide sequence analysis of VP7 gene of rotavirus isolated from infants in Korea could be used for identification, serotying and determination of novel or unusual strains of rotaviruses.

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Anti-herpes Activity of Vinegar-processed Daphne genkwa Flos Via Enhancement of Natural Killer Cell Activity

  • Uyangaa, Erdenebileg;Choi, Jin Young;Ryu, Hyung Won;Oh, Sei-Ryang;Eo, Seong Kug
    • IMMUNE NETWORK
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    • v.15 no.2
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    • pp.91-99
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    • 2015
  • Herpes simplex virus (HSV) is a common causative agent of genital ulceration and can lead to subsequent neurological disease in some cases. Here, using a genital infection model, we tested the efficacy of vinegar-processed flos of Daphne genkwa (vp-genkwa) to modulate vaginal inflammation caused by HSV-1 infection. Our data revealed that treatment with optimal doses of vp-genkwa after, but not before, HSV-1 infection provided enhanced resistance against HSV-1 infection, as corroborated by reduced mortality and clinical signs. Consistent with these results, treatment with vp-genkwa after HSV-1 infection reduced viral replication in the vaginal tract. Furthermore, somewhat intriguingly, treatment of vp-genkwa after HSV-1 infection increased the frequency and absolute number of $CD3^-NK1.1^+NKp46^+$ natural killer (NK) cells producing interferon (IFN)-${\gamma}$ and granyzme B, which indicates that vp-genkwa treatment induces the activation of NK cells. Supportively, secreted IFN-${\gamma}$ was detected at an increased level in vaginal lavages of mice treated with vp-genkwa after HSV-1 infection. These results indicate that enhanced resistance to HSV-1 infection by treatment with vp-genkwa is associated with NK cell activation. Therefore, our data provide a valuable insight into the use of vp-genkwa to control clinical severity in HSV infection through NK cell activation.