• The Plant Pathology Journal
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• v.23 no.4
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• pp.227-232
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• 2007

In the plant virus world, there are six genera of plant viruses with dsDNA genomes and six genera with ssDNA (Fauquet et al., 2005). The dsDNA viruses are comprised of 4 genera in the Caulimoviridae, the genus Badnavirus and the genus Tungrovirus. The ssDNA viruses are comprised of four genera in Geminiviridae, and the two genera Nanovirus and Babuvirus in the Nanoviridae. The genera Babuvirus and Badnavirus are not well studied in Asia. However, we recognized the significance of two species, Banana bunchy top virus (BBTV) in the genus Babuvirus and Banana streak virus (BSV) in the genus Badnavirus, during the survey of banana viruses in Asia. Their main characters will be introduced in this mini-review.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.285-291
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• 2007

According to the Baltimore Scheme, viruses are classified into 6 main classes based on their replication and coding strategies. Except for some small DNA viruses, most viruses code for their own polymerases: DNA-dependent DNA, RNA-dependent RNA and RNA-dependent DNA polymerases, all of which contain 4 common motifs. We undertook a phylogenetic study to establish the relationship between the Baltimore Scheme and viral polymerases. Amino acid sequence data sets of viral polymerases were taken from NCBI GenBank, and a multiple alignment was performed with CLUSTAL X program. Phylogenetic trees of viral polymerases constructed from the distance matrices were generally consistent with Baltimore Scheme with some minor exceptions. Interestingly, negative RNA viruses (Class V) could be further divided into 2 subgroups with segmented and non-segmented genomes. Thus, Baltimore Scheme for viral taxonomy could be supported by phylogenetic analysis based on the amino acid sequences of viral polymerases.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.186-191
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• 2002

The usefulness of oligonucleotide DNA chip was evaluated for detection and primary level identification of major waterborne viruses in environmental samples. The enteric waterborne viruses included enterovirus, adenovirus, and rotavirus. Total intracellular RNA of 10 BGM cell plates showing virus-specific cytopathic effects was extracted at the third day after inoculation. The intracellular RNA was then subjected to either enterovirus-specific RT-PCR followed by sequencing analysis, or the DNA chip. Seven out of 10 positive samples in cell culture were positive but the other three sample were turned out to be negative by both RT-PCR and DNA chip analyses. Nucleotide sequencing results and the DNA chip hybridization results of the RT-PCR product were in complete agreement in the identification of the 7 positive samples as enteroviruses. Using the DNA chip, it took only 3∼4 hr to complete detection and primary level identification of target viruses and additional procedures such as gel electrophoresis or nucleotide sequencing were not necessary. We believe that the DNA chip system can be employed as a highly effective and new detection methodology for environmental viruses.

• The Plant Pathology Journal
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• v.17 no.3
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• pp.164-168
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• 2001

Rice black-streaked dwarf virus (RBSDV) and Maize rough dwarf virus (MRDV) are closely related viruses. Since the two viruses produce identical symptoms on maize, barley, and wheat, diagnosis of infected plants is difficult. Previously, we reported that partial cDNA clones of RBSDV S5 and S6 from the Japanese isolate (RBSDV-H) have lower sequence homology to MRDV than do cDNA clones from other genomic segments. In order to test whether cDNA clones of RBSDV-H S5 and S6 can be used for molecular diagnosis, RBSDV field isolates from Korea and from Hokkaido, Japan were tested in dot blot hybridizations probed with RBSDV-H S5 and S6 cDNA colnes. Hybridization with these probes was more intense against the RBSDV genome than against the MRDV genome. Therefore, RBSDV-H S5 and S6 cDNA clones can be used to differentiate between the two viruses. Furthermore, RBSDV-H S5 and S6 clones reacted more strongly against the viruses from stunted maize plants from Korean fields than to MRDV, indicating that RBSDV may be the causal disease agent in maize plants in Korea.

• The Plant Pathology Journal
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• v.21 no.1
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• pp.13-20
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• 2005

Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.

• Research in Plant Disease
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• v.30 no.3
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• pp.304-311
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• 2024

There are concerns about the introduction and spread of plant pests and pathogens with globalization and climate change. As commercial control agents have not been developed for plant viruses, it is important to prevent virus spread. In this study, we developed a multiplex polymerase chain reaction (PCR) detection method to rapidly diagnose and control three DNA (papaya golden mosaic virus, Lindernia anagallis yellow vein virus, and melon chlorotic leaf curl virus) and two RNA (papaya leaf distortion mosaic virus and lettuce chlorosis virus) viruses that infect papaya. Specific primer sets were designed for the virus coat protein. Performing PCR, clear bands were observed with no non-specific reaction. Our multiplex PCR method can simultaneously detect small amounts of DNA/RNA to diagnose five viruses infecting papaya and prevent the spread of the virus.

• Applied Biological Chemistry
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• v.36 no.4
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• pp.315-317
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• 1993

To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses (GV), virus particles were isolated from field-grown garlic leaves and RNA genome was isolated from them. It was used for constructing cDNA library for GV. Several cDNA clones for GV were isolated and classified into 4 different groups on the basis of cross Southern hybridization. Northern blot analysis of GV RNA with one of these cDNA clones shows that the clone is a cDNA for GV RNA.

• Korean journal of applied entomology
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• v.30 no.2
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• pp.138-143
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• 1991

This study was carried out to acquire some basic biochemical informations on the granulosis virus (GV) DNA of Pieris rapae and Pieris brassicae. The thermal denaturation temperature (Tm) and G+C content of the DNA of the viruses were $83.7^{\circ}C$ and 35.5% for P. rapae GV, $84.0^{\circ}C$ and 35.9% for P. brassicae GV, respectively. There were some differences in the DNA fragmentation patterns of the two GV's produced by digestion with restriction endonucleases such as EcoR I , BamH I and Hind m . The homololgy between the two DNAs was caculated to be 97.0%. The size of the genome was estimated to be 103 kbp for P. rapae GV and 108 kbp for P. brassicae GV.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1164-1169
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• 2008

We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. RT-PCR products with unique sizes characteristic of each subtype were amplified by multiplex RT-PCRs, and sequence analysis of each amplicon was demonstrated to be specific for each subtype with 24 reference viruses. The specificity was demonstrated further with DNA or cDNA templates from 7 viruses, 5 bacteria, and 50 influenza A virus-negative specimens. Furthermore, the assays could detect and subtype up to $10^5$ dilution of each of the reference viruses that had an original infectivity titer of $10^6\;EID_{50}/ml$. Of 188 virus isolates, the multiplex RT-PCR results agreed completely with individual RT-PCR subtyping results and with results obtained from virus isolations. Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes of influenza viruses in one host. Therefore, these methods could facilitate rapid and accurate subtyping of influenza A viruses directly from field specimens.

• BMB Reports
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• v.28 no.3
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• pp.216-220
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• 1995

cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.