• Molecules and Cells
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• v.42 no.1
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• pp.67-78
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• 2019

Methylation of HBV cccDNA has been detected in vivo and in vitro; however, the mechanism and its effects on HBV replication remain unclear. HBx derived from a 1.2-mer HBV replicon upregulated protein levels and enzyme activities of DNA methyltransferase 1 (DNMT1), 3a, and 3b, resulting in methylation of the negative regulatory region (NRE) in cccDNA, while none of these effects were observed with an HBx-null mutant. The HBx-positive HBV cccDNA expressed higher levels of HBc and produced about 4-fold higher levels of HBV particles than those from the HBx-null counterpart. For these effects, HBx interrupted the action of NRE binding protein via methylation of the C-1619 within NRE, resulting in activation of the core promoter. Treatment with 5-Aza-2′dC or DNMT1 knock-down drastically impaired the ability of HBx to activate the core promoter and stimulate HBV replication in 1.2-mer HBV replicon and in vitro infection systems, indicating the positive role of HBx-mediated cccDNA methylation in HBV replication.

• Microbiology and Biotechnology Letters
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• v.15 no.5
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• pp.319-327
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• 1987

In order to develop useful plasmid vectors for Zymomonas cells, attempts were made to isolate natural plasmids from Z. mobilis ATCC10988. Among a few plasmids isolated, a small plasmid of 3.9 Kb size was chosen and designated as pZM3. By introducing the replication origin of pZM3 into pBR325, a hybrid plasmid vector of 8.4 Kb size, pHZ22, was constructed. This vector contained chloramphenicol resistant gene as a selectable marker and proved to be conjugally transmissible and stably maintained in Z. mobilis. Tetracycline resistant gene was isolated from RP4 and introduced into pHZ22 to make a new vector called pHZT224 of 10.7 Kb size. Through n series of experiments, it was evident that these plasmid vectors containing selectable markers of chloramphenicol and tetracycline resistance were shuttle vectors functional in Z. mobilis as well as E. coli.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1229-1239
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• 2005

Human immunodeficiency virus-like particles (HIVVLPs) with native conformations similar to that of the wild-type virion could be valid candidates for vaccine development. To this end, we used a Semliki Forest Virus (SFV) expression system to produce HIV- VLPs containing high quantities of native envelope proteins. Here, we described a single SFV replicon containing the HIV gagpol and env genes under the control of separate subgenomic promoters. Mature VLPs incorporating the Gag and Env proteins were detected in the supernatant of replicon-expressing cells by Western blot analysis. The HIV-VLPs showed the expected molecular density (1.14-1.18 g/ml) on a $20-60\%$ sucrose gradient; the particles were 100-120 nm in diameter and Env proteins were observed on their surfaces by immunogold electron microscopy. RT-PCR analysis of VLP-associated RNAs in mature HIV-VLPs revealed two SF V-derived RNA species (full-length and subgenomic). Immunization studies in Balb/c mice showed that these HIV-VLPs were capable of inducing both HIV-specific antibodies and cell-mediated immune responses. Taken together, our results indicate that the SFV replicon system is useful for the production of HIV-VLPs, which may be valuable candidates for an HIV vaccine.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.155-160
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• 2001

We have generated a novel baculovirus genome which can be maintained in Escherichia coli that facilitates the rapid and efficient generation of recombinant baculovirus expression vectors. To make $Occ^{+}$ recombinant expression vectors, polyhedrin gene under the control of p10 promoter was inserted to bAcGOZA and this genome was designated bApGOZA. As in bAcGOZA, bApGOZA lacks a portion of the essential ORF1629 gene, but includes a mini-F replicon and selectable kanamycin-resistance marker, This occasion-producing activity of bApGOZA can be used very conveniently for its oral infectivity to insect larvae in mass production of foreign protein and insecticides.

• Food Science and Industry
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• v.50 no.1
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• pp.67-73
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• 2017

Control of food pathogens is critical in food safety field. Norovirus is one of the major causes of gastroenteritis and food poisoning worldwide, however, currently, there is not a vaccine or a specific drug available for its treatment. There are several methods to inactivate norovirus during food processing by chemical and physical treatments, however, the use of natural substance has been suggested as an optional strategy due to their safety and consumer preference. In this study supported by Ministry of Food and Drug Safety in Korea, we identified novel plant-derived substances with significant anti-norovirus activities. The aim of this project was to determine the antiviral activity of a wide range of natural substances, including plant-derived extracts and essential oils, using a norovirus surrogate system, human norovirus replicon-bearing cells, and mouse in vivo experiments. During the activity screening test, we identified novel anti-norovirus substances or oils using plaque assay with MNV-1. Six selected substances were formulated into an optimum mixture and used as an ingredient for salad sauce of which anti-novovirus activity was confirmed(pending for patent and paper submission). The potential application of selected natural substances as a metal surface sanitizer was also tested. Interestingly, the mixture of selected natural compounds showed a significant inhibitory effect against norovirus. These results suggest that these substances may be used as food ingredient with anti-norovirus antivity or components for surface sanitizers to prevent norovirus contamination.

• Korean Journal of Environmental Agriculture
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• v.1 no.1
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• pp.48-52
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• 1982

Methods for the measurement of aquatic bacteria can be divided into two groups. The first group of these methods is based on the 'replicon' concept that live bacterial cells, when diluted and transferred to a suitable medium, produce colonies. These methods distinguish living from dead bacteria, but they massively underestimate bacterial numbers. The second group of enumeration methods uses visual counting technique using specific apparatus such as a microscope. These methods are generally direct and simple, but it is very hard to distinguish between live and dead bacteria and between small particle and bacteria. Recently developed technique in staining methods has provided a reliable method of visual determination of aquatic bacteria. This uses epifluorescence microscopy to measure the total bacterial population. In order to present the fluorescence microscopy as a new methodology for the determination of bacterial numbers in water supplies, data were obtained from chlorine and monochloramine doses added to samples. Total counts by fluorescence microscopy were compared with standard plate count method. The total number of bacteria in water supplies can be determined with fluorescence microscopy. This technique allows better resolution of small bacteria and differentiation of particle from bacteria. Chloramine was found to persist longer in natural waters and prevent bacterial regrowth.