• Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.185-187
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• 2005

We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

• Proceedings of the Korean Society of Life Science Conference
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• 2007.10a
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• pp.56.2-56.2
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• 2007

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• The Plant Pathology Journal
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• v.15 no.4
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• pp.247-250
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• 1999

Wasabies showing mosaic symptoms were collected and extracted for virus purification. Tobacco mosaic virus (TMV) was identified as causal agent by electron microscopy and nucleic acid and coat protein analyses. TMV strains were determined by enzyme-linked immunosorbent assay (ELISA). TMV was identified as W and C strain in wasabi. The results of host reaction indicated that this virus induced local lesions on Nicotiana tabacum cv. Bright Yellow and N. glutinosa, leaf spots on Chenopodium amaranticolor and mosaic symptoms on wasabi. Rot shape virus particles were observed and was about 300 nm in length. About 6.5 kb single RNA molecule was observed from extracted viral RNA sample and 26 KDa coat protein was detected in denatured acrylamide gel. Infection ratio of TMV was 8% for the first cultivation year, but was 22% for the second year when TMV-W antiserum was used. The results of this experiment showed that infection ratios of both TMV-W and TMV-C strains were higher compared to that of TMV-P strain.

• The Plant Pathology Journal
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• v.20 no.4
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• pp.313-315
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• 2004

The coat protein gene of Korean isolate of Ricer stripe virus (RSV-Kr) was cloned and its nucleotide sequence was determined. Total RNA was extracted from infected leaves and RSV viral RNA was detected by using RT-PCR with specific primer of coat protein gene. The result of RT-PCR showed a specific band. Purified RT-PCR products of coat protein gene were ligated into the pGEM-T Easy plasmid vector and cloned cDNA was obtained for nucleotide sequence determination. Coat protein gene of RSV-Kr consisted of 969 bp long encoding a protein of 322 amino acids. RSV-Kr showed 94%-99% sequence identities to that of Japanese- and Chinese isolates.

• The Plant Pathology Journal
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• v.22 no.2
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• pp.164-167
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• 2006

An immunocapture reverse transcription-polymerase chain reaction (IC/RT-PCR) was developed for simultaneous detection of two pepper-infecting RNA viruses, Pepper mud mottle virus (PMMoV) and Tobacco mild green mosaic virus (TMGMV). The assay could be performed in a single tube for simultaneous and sensitive detection of these tobamoviruses. This detection system revealed thousand-fold increase in detection sensitivity compare to ELISA. This method could save time and reagent cost compare to common RT-PCR which needs several reactions and several procedures of viral RNA extractions for the same number of samples.

• Korean Journal Plant Pathology
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• v.11 no.1
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• pp.87-90
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• 1995

Complementary DNA of the movement protein (MP) gene of tobacco mosaic virus Korean pepper strain (TMV-KP) was synthesized from purified TMV-KP RNA by using the reverse transcription and polymerase chain reaction (PCR) system. The synthesized double stranded cDNA was cloned into the plasmid pUC9 and transformed into Escherichia coli JM110. The movement protein gene of TMV-KP of the selected clones was subjected to sequence analysis by Sanger's dideoxy chain termination method. The complete sequence of viral MP gene from TMV-KP strain was 807 nucleotides long. The nucleotide of MP gene from TMV-KP has thirteen and two nucleotide differences from TMV vulgarae (TMV-OM) and Korean (TMV-K) strains, respectively. Thus, the nucleotide sequence of TMV-KP MP gene showed higher homology of 99% with that of TMV-K MP gene.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.315-323
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• 2011

Soybean plants infected with Bean pod mottle virus (BPMV) develop acute symptoms that usually decrease in severity over time. In other plant-virus interactions, this type of symptom recovery has been associated with degradation of viral RNAs by RNA silencing, which is accompanied by the accumulation of virus-derived small interfering RNAs (siRNAs). In this study, changes in the accumulation of BPMV siRNAs were investigated in soybean plants infected with BPMV alone, or infected with both BPMV and Soybean mosaic virus (SMV) and in transgenic soybean plants expressing SMV helper component-protease (HC-Pro). In many potyviruses, HC-Pro is a potent suppressor of RNA silencing. In plants infected with BPMV alone, accumulation of siRNAs was positively correlated with symptom severity and accumulation of BPMV genomic RNAs. Plants infected with both BPMV and SMV and BPMV-infected transgenic soybean plants expressing SMV HC-Pro exhibited severe symptoms characteristic of BPMVSMV synergism, and showed enhanced accumulation of BPMV RNAs and siRNAs compared to plants infected with BPMV alone and nontransgenic plants. Likewise, SMV HC-Pro enhanced the accumulation of siRNAs produced from a silenced green fluorescent protein gene in transient expression assays, while the P19 silencing suppressor of Tomato bushy stunt virus did not. Consistent with the modes of action of HC-Pro in other systems, which have shown that HC-Pro suppresses RNA silencing by preventing the unwinding of duplex siRNAs and inhibiting siRNA methylation, these studies showed that SMV HC-Pro interfered with the activities of RNA-induced silencing complexes, but not the activities of Dicer-like enzymes in antiviral defenses.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.130-138
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• 2010

Orchid fleck virus (OFV) is an unassigned plant virus in the family Rhabdoviridae. OFV was isolated from Cymbidium sp. showing oval necrotic lesions on their leaves in Korea, and designated as OFV-NHHS1. The complete nucleotide sequence of the RNA1 (6,413 nt) (GenBank accession no. AB516442) and RNA2 (6,001 nt) (GenBank accession no. AB516441) was determined in this study. RNA1 and RNA2 contained five and one ORF respectively. RNA1 encodes nucleocapsid (N) of 49 kDa, ORF2 of 26 kDa, ORF3 of 38 kDa, ORF4 of 20 kDa and glycoprotein (G) of 61 kDa proteins, whereas RNA2 encodes a single polymerase of 212 kDa. OFV-NHHS1 shared extremely high similarity of 98.6-100% and 98.9-99.6% in nucleotidle and amino acid sequences with a Japanese isolate, OFV-so, respectively. However, the N, G and L of OFV-NHHS1 revealed 6.9-19.3%, 7.3-12.0%, and 13.4-26.6% identities to those of 29 Rhabdoviruses, respectively. To survey the infection rate of OFV in commercial orchids in Korea, 51 Cymbidium sp., 10 Phalaenopsis sp., 22 Oncidium sp. and 21 Dendrobium sp. plants that showed typical viral symptoms were collected. RT-PCR with specific primers for detection of Cymbidium mosaic virus (CymMV), ORSV and OFV showed high infection rate by ORSV alone and double infection by ORSV and CymMV. One of the orchids tested was infected with OFV. This is the first report of the complete nucleotide sequences of OFV isolated in Korea.

• The Plant Pathology Journal
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• v.36 no.6
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• pp.643-650
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• 2020

Korean ginseng (Panax ginseng) is a dicotyledonous, medicinal, perennial plant belonging to the genus Panax of the family Araliaceae. We investigated the occurrence and incidence of plant viruses in Panax ginseng in Korea. A total of 656 leaf samples were combined into one and total RNA was extracted from the polled sample, using RNA sequencing (RNA-Seq), a metatranscriptome analysis of the plant virome was conducted. The virus present in Panax ginseng was confirmed by reverse transcription polymerase chain reaction (RT-PCR) assay using virus-specific primers. In RNA-Seq data analysis, the multiplication protein of four viral contigs including Aristotelia chilensis virus 1 (AcV1), Turnip mosaic virus (TuMV), Watermelon mosaic virus (WMV), and Tobamovirus multiplication protein were discovered. From our metatranscriptome analysis and RT-PCR assay, TuMV and WMV were detected, whereas the three viruses reported in China such as tomato yellow leaf curl China virus; panax notoginseng virus A; and panax virus Y were not found in this study. The distribution of domestic ginseng viruses seems different from that recorded in China. Overall, this is the first plant virome analysis of Panax ginseng in Korea.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.277.1-277.1
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• 2013

Molecular diagnostics consists of three processes, which are a sample pretreatment, a nucleic acid amplification, and an amplicon detection. Among three components, sample pretreatment is an important process in that it can increase the limit of detection by purifying nucleic acid in biological sample from contaminants that may interfere with the downstream genetic analysis such as nucleic acid amplification and detection. To achieve point-of-care virus detection system, the sample pretreatment process needs to be simple, rapid, and automatic. However, the commercial RNA extraction kits such as Rneasy (Qiagen) or MagnaPure (Roche) kit are highly labor-intensive and time-consuming due to numerous manual steps, and so it is not adequate for the on-site sample preparation. Herein, we have developed a rotary microfluidic system to extract and purify the RNA without necessity of external mechanical syringe pumps to allow flow control using microfluidic technology. We designed three reservoirs for sample, washing buffer, and elution buffer which were connected with different dimensional microfluidic channels. By controlling RPM, we could dispense a RNA sample solution, a washing buffer, and an elution buffer successively, so that the RNA was captured in the sol-gel solid phase, purified, and eluted in the downstream. Such a novel rotary sample preparation system eliminates some complicated hardwares and human intervention providing the opportunity to construct a fully integrated genetic analysis microsystem.