• KSBB Journal
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• v.11 no.4
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• pp.438-444
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• 1996

Using recombinant Vaccinia virus(vSC8) that express ${\beta}$-galactosidase, a model heterologous protein, conditions for virus and protein production were investigated in tissue culture flask. As host animal cells HeLa and HeLa S3 were used. It was demonstrated that cells infected during the exponential growth phase gave higher protein yield than those infected during the stationary growth phase and calf serum concentration after virus infection did not significantly alter protein yield. Pretreatment of cell layer with hypotonic solution enhanced the virus infectivity. Optimum cell growth and recombinant protein production was achieved at $37^{\circ}C$. But, during 2 hours of virus infection period incubation temperature must be lowered to 20∼$30^{\circ}C$ for maximum recombinant protein yield. To enhance virus replication, the effects of adrenal glucocorticoid hormone (Dexamethasone) and silkworm hemolymph were evaluated. Only dexamethasone increased about 20% of ${\beta}$-galactosidase yield in HeLa S3 cells when added with 10-7∼10-5M concentration 24 hours before infection.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2018.10a
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• pp.444-447
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• 2018

Baculovirus expression systems have many known advantages including fast and cost-effective methods to generate large amounts of recombinant proteins in comparison to bacterial expression systems, particularly those requiring complex post-translational modifications. Especially, recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. In this study, baculoviral vectors which were reconstructed from pcDNA3.1 vector, were recombined with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector.

• Journal of Sericultural and Entomological Science
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• v.27 no.2
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• pp.20-27
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• 1985

This study was carried out to investigate the histological changes after the infection of nuclear and cytoplasmic polyhedrosis viruses(NPV, CPV) and the resistance to the viruses in various varieties of the silkworm fed on artificial diet. The results obtained were as follows; Among four varieties of silkworm tested, Jam 107$\times$Jam 108 was more resistant than the other varieties tested and Jam 119$\times$Jam 120 was the most resistant to CPV. In case of peroral infection with NPV, Jam 107$\times$Jam 108 showed lower mortality than the remained varieties in low concentration (104/ml). However, all varieties showed high mortality as the concentration of viruses was increased. With infection of CPV, the varieties showed high mortality at the concentration of 107 and 108/ml, while Jam 119$\times$Jam 120 showed the lowest mortality at virus concentration of 104/ml. The fat bodies, epidermal cells and tracheal epithelial cells showed high susceptibility to NPV to break the cells completely and liberate the debris to the body cavity. The CPV infected only the cylindrical cells of mid-gut and formed polyhedrons. In some cells, CPV was liberated to gastral cavity. In the electrophoretic pattern of hemolymph protein of silkworm larvae infected with NPV, bands were dimmed and disappeared as symptom aggravated after infection. Electrophoretic pattern of homolymph proteins of silkworm larvae infected with CPV showed no numerical difference at the later stage of infection, and one or two bands was observed along with lowering the concentrations.

• Korean Journal of Poultry Science
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• v.38 no.2
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• pp.97-104
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• 2011

We conducted a 10-month (March to October 2009) surveillance of Newcastle disease virus (NDV) in 13 slaughterhouses in Korea. NDV was isolated in 13.0%, 13.3%, 16.0%, and 10.8% of chicken farms, transport vehicles, hang rooms, and chilling water, respectively. Of NDV isolates from slaughterhouses, 37% were isolated in July. All NDV isolates were determined to be lentogenic viruses by RT-PCR-based pathotyping, and all NDV isolates had the $^{112}GKQGR/L^{117}$ motif at the cleavage site of the F protein. Phylogenetic analysis based on the hypervariable region of the F protein gene classified all 25 NDV isolates examined into genotype I within class II. Of these, 24 were clustered together with the NDV V4 strain, while the remaining isolate was placed in the cluster belonging to the NDV Ulster 2C strain. Our results indicate that lentogenic NDV was a high-frequency contaminant in the serial process of ranging live birds to slaughtering at slaughterhouses.

• Research in Plant Disease
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• v.21 no.2
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• pp.99-102
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• 2015

Cucumber mosaic virus (CMV) is one of the most destructive viruses in chilli pepper. An isolate of CMV was obtained from the chilli pepper cv. Chungyang showing top necrosis symptom in 2013 and designated as CMV-GTN. CMV-GTN was compared with the well-characterized isolate, CMV-Ca-P1, by investigating their amino acid sequences of the coat protein (CP) and biological reactions in several host plants. The CP of CMV-Ca-P1 composed of 217 amino acids but that of CMV-GTN composed of 218 amino acids by including additional valine in the $57^{th}$ amino acid position. Amino acid sequence similarity of the CP gene among CMV-GTN and other CMV isolates recorded in the GeneBank database ranged from 96% to 99%. CMV-GTN was selected as a representative isolate to screen the resistance pepper cultivars to CMV because it was highly pathogenic to tomatoes and peppers upon biological assays. The virulence of CMV-GTN was tested on 135 pepper cultivars which has been bred in Korea and compared with that of CMV-Ca-P1. Only the cv. Premium was resistant and three cvs. Hot star, Kaiser, and Good choice were moderately resistant to CMV-GTN, whereas two cvs. Baerotta and Kaiser were resistant to CMV-Ca-P1.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.168-176
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• 2010

Acellular dermal matrix (ADM), produced by decellularization from human cadaveric skin, has been used for various biomedical applications. A manufacturing process for ADM ($SureDerm^{TM}$) using tri-n-butyl phospahate (TnBP) and deoxycholic acids as the decellularization solution has been developed. The manufacturing process for $SureDerm^{TM}$ has 70% ethanol treatment and ethylene oxide gas sterilization for inactivating infectious microorganisms. The purpose of this study was to examine the efficacy of the 70% ethanol treatment, decellularization process using 0.1% TnBP and 2% deoxycholic acids, and EO gas sterilization process in the inactivation of viruses. A variety of experimental model viruses for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), and porcine parvovirus (PPV) were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment. However HAV and PPV showed high resistance to 70% ethanol treatment with the log reduction factors of 1.85 and 1.15, respectively. HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by decellularization process. All the viruses tested were completely inactivated to undetectable levels by EO gas treatment. The cumulative log reduction factors of HIV-1, BHV, BVDV, HAV, and PPV were $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, and $\geq7.18$, respectively. These results indicate that the production process for $SureDerm^{TM}$ has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.161-166
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• 1998

To study the usefulness of Se301 cells, which is originated from Spodoptera exigua and has susceptibility to the Autographa californica NPV (AcNPV), as a host for the AcNPV-based expression vector system, we compared the characteristics of AcNPV in Se301 and Sf-21 cells. The symptom by viral infection was similar in both of cells, but the ratio of polyhedra released from the cell was higher in Se301 cells than in Sf-21 cells. The overall PIB productivity of AcNPV was similar in both cells but the size of polyhedra was larger in Se301 cells. While the polyhedrin expression efficiency was about 2.4 times higher in Se301 cells than in Sf-21 cells, the viral growth was higher in Sf-21 cells. These results suggested that Se301 cell is very useful in the AcNPV-based expression system as a host.

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.175-179
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• 1999

In order to find less toxic antiviral agents from Basidiomycetes, EA, the water soluble substance, was prepared from the carpophores of Elfvingia applanata (Pers.) Karst. Antiviral activity of EA against vesicular stomatitis virus [New Jersey serotype, VSV(NJ)] was examined in Vero cells using plaque reduction assay in vitro. And the combined antiviral effects of EA with interferon (IFN) alpha and gamma were examined on the multiplication of VSV(NJ). EA caused a concentration-dependent reduction in the plaque formation of VSV(NJ) with 50% effective concentration $(EC_{50})$ of 2.10 mg/ml. The results of combination assay were evaluated by the combination index (CI) that was analysed by the multiple drug effect analysis. The combination of EA with IFN alpha showed more potent effect with CI values of $0.87{\sim}1.59$ for 50%, 70% and 90% effective levels than that with INF gamma with CI values of $1.05{\sim}2.03$.

• The Journal of Korean Society of Virology
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• v.28 no.4
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• pp.383-391
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• 1998

A murine monoclonal antibody (mAb) specific for the envelope glycoprotein gp120 of human immunodeficiency virus type-I (HIV -1) was chemically coupled to pokeweed antiviral protein (PAP) from Phytolacca americana. The immunotoxin was purified by FPLC using S200 colum. The purified immunotoxin efficiently bound to HIV-infected T cells as evidenced by fluorescenceactivated cell sorter analysis. The immunotoxin selectively killed human T lymphoid lines infected with $HIV-1_{IIIB}$ at less than 250 pM of the immunotoxin cells, while PAP or mAb alone did not have any significant effect on infected cells. The uninfected control T cell lines were not affected. Human cells infected with HIV-2 or other HIV-1 strains were not killed, suggesting that the killing depends completely on the antibody used for coupling. These in vitro results suggest that the PAP-mAb conjugate may be used to selectively remove cells expressing viral antigens from individuals infected with HIV.

• Journal of Plant Biology
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• v.38 no.3
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• pp.267-274
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• 1995

Based upon the nucleotide sequence of As strain of cucumber mosaic virus (CMV-As0 RNA4, coat protein (CP) gene was selected for the design of oligonucleotide primers of polymerase chain reaction (PCR) for detection and identification of the virus. Reverse transcription and polymerase chain reaction (RT-PCR) was performed with a set of 18-mer CMV CP-specific primers to amplify a 671 bp fragment from crude nucleic acid extracts of virus-infected leaf tissues as well as purified viral RNAs. The minimum concentrations of template viral RNA and crude nucleic acids from infected tobacco tissue required to detect the virus were 1.0 fg and 1:65,536 (w/v), respectively. No PCR product was obtained when potato virus Y-VN RNA or extracts of healthy plants were used as templates in RT-PCR using the same primers. The RT-PCR detected CMV-Y strain as well as CMV-As strain. Restriction analysis of the two individual PCR amplified DNA fragments from CMV-As and CMV-Y strains showed distinct polymorphic patterns. PCR product from CMV-As has a single recognition site for EcoRI and EcoRV, respectively, and the product from CMV-Y has no site for EcoRI or EcoRV but only one site for HindIII. The RT-PCR was able to detect the virus in the tissues of infected pepper, tomato and Chinese cabbage plants.