Publications by authors named "Eui-Joon Kil"

32 Publications

Adaptation and Codon-Usage Preference of Apple and Pear-Infecting Apple Stem Grooving Viruses.

Microorganisms 2021 May 21;9(6). Epub 2021 May 21.

Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Korea.

Apple stem grooving virus (ASGV; genus ) is an economically important virus. It has an approx. 6.5 kb, monopartite, linear, positive-sense, single-stranded RNA genome. The present study includes identification of 24 isolates-13 isolates from apple ( L.) and 11 isolates from pear ( L.)-from different agricultural fields in South Korea. The coat protein (CP) gene of the corresponding 23 isolates were amplified, sequenced, and analyzed. The CP sequences showed phylogenetic separation based on their host species, and not on the geography, indicating host adaptation. Further analysis showed that the ASGV isolated in this study followed host adaptation influenced and preferred by the host codon-usage.
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http://dx.doi.org/10.3390/microorganisms9061111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8223792PMC
May 2021

Transcriptome Assembly of Two Fern Species Identifies Enzymes Required for Two Upstream Pathways of Phytoecdysteroids.

Int J Mol Sci 2021 Feb 19;22(4). Epub 2021 Feb 19.

Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Korea.

species produce a high amount of phytoecdysteroids (PEs), which are widely used in traditional medicine in the Pacific islands. The PEs in two different species, (MP) and (MS), were examined using high-performance liquid chromatography (HPLC). In particular, MS produces a high amount of 20-hydroxyecdysone, which is the main active compound in PEs. To identify genes for PE biosynthesis, we generated reference transcriptomes from sterile frond tissues using the NovaSeq 6000 system. De novo transcriptome assembly after deleting contaminants resulted in 57,252 and 54,618 clean transcripts for MP and MS, respectively. The clean transcripts for each species were annotated according to gene ontology terms, UniProt pathways, and the clusters of the orthologous group protein database using the MEGAN6 and Sma3s programs. In total, 1852 and 1980 transcription factors were identified for MP and MS, respectively. We obtained transcripts encoding for 38 and 32 enzymes for MP and MS, respectively, potentially involved in mevalonate and sterol biosynthetic pathways, which produce precursors for PE biosynthesis. Phylogenetic analyses revealed many redundant and unique enzymes between the two species. Overall, this study provides two reference transcriptomes that might be useful for further studies regarding PE biosynthesis in species.
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http://dx.doi.org/10.3390/ijms22042085DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7923240PMC
February 2021

Nanovirus Disease Complexes: An Emerging Threat in the Modern Era.

Front Plant Sci 2020 19;11:558403. Epub 2020 Nov 19.

Department of Integrative Biotechnology, Sungkyunkwan University, Suwon, South Korea.

Multipartite viruses package their genomic segments independently and mainly infect plants; few target animals. is a family of multipartite single-stranded DNA plant viruses that individually encapsidate single-stranded DNAs of approximately 1 kb and transmit them through aphids without replication in the aphid vectors, thereby causing important diseases of leguminous crops and banana. Significant findings regarding nanoviruses have recently been made on important features, such as their multicellular way of life, the transmission of distinct encapsidated genome segments through the vector body, evolutionary ambiguities, mode of infection, host range and geographical distribution. This review deals with all the above-mentioned features in view of recent advances with special emphasis on the emergence of new species and recognition of new host range of nanoviruses and aims to shed light on the evolutionary linkages, the potentially devastating impact on the world economy, and the future challenges imposed by nanoviruses.
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http://dx.doi.org/10.3389/fpls.2020.558403DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7710663PMC
November 2020

Rapid diagnosis of two marine viruses, red sea bream iridovirus and viral hemorrhagic septicemia virus by PCR combined with lateral flow assay.

Virusdisease 2020 Sep 31;31(3):251-256. Epub 2020 Mar 31.

College of Biotechnology and Bioengineering, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 16419 Korea.

Red sea bream iridovirus (RSIV) and Viral hemorrhagic septicemia virus (VHSV are the most important viral marine pathogens in South Korea because RSIV and VHSV infect and cause high mortality rates in major fish species such as and . These viruses can be transmitted both vertically and horizontally, and early diagnosis is imperative. In this research, RSIV and VHSV viral genomes are detected by PCR-lateral flow assay (LFA). PCR-LFA is sensitive, capable of detecting a viral genome at a concentration of 2-200 fg/µL. Development of this detection method is very meaningful because LFA is simple, requiring a minimum of personnel training to perform. Additionally, LFA requires less time than other detection methods and can be an immediate detection tool that is indispensable in preventing rapid viral spread.
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http://dx.doi.org/10.1007/s13337-020-00577-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459051PMC
September 2020

Development of novel detection system for sweet potato leaf curl virus using recombinant scFv.

Sci Rep 2020 05 15;10(1):8039. Epub 2020 May 15.

College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon, 16419, Republic of Korea.

Sweet potato leaf curl virus (SPLCV) causes yield losses in sweet potato cultivation. Diagnostic techniques such as serological detection have been developed because these plant viruses are difficult to treat. Serological assays have been used extensively with recombinant antibodies such as whole immunoglobulin or single-chain variable fragments (scFv). An scFv consists of variable heavy (V) and variable light (V) chains joined with a short, flexible peptide linker. An scFv can serve as a diagnostic application using various combinations of variable chains. Two SPLCV-specific scFv clones, F7 and G7, were screened by bio-panning process with a yeast cell which expressed coat protein (CP) of SPLCV. The scFv genes were subcloned and expressed in Escherichia coli. The binding affinity and characteristics of the expressed proteins were confirmed by enzyme-linked immunosorbent assay using SPLCV-infected plant leaves. Virus-specific scFv selection by a combination of yeast-surface display and scFv-phage display can be applied to detection of any virus.
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http://dx.doi.org/10.1038/s41598-020-64996-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7228925PMC
May 2020

Seed Transmission of Tomato Leaf Curl New Delhi Virus from Zucchini Squash in Italy.

Plants (Basel) 2020 Apr 29;9(5). Epub 2020 Apr 29.

College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 16419, Korea.

(ToLCNDV) is a bipartite begomovirus affecting tomato cultivation on the Indian subcontinent. Recently, however, a new strain of the virus, named ToLCNDV-ES, has spread to Mediterranean countries such as Spain, Italy, and Tunisia, and occurred in Cucurbita crops, causing economic damage. Although ToLCNDV is spread by the sweet potato whitefly (), like other begomoviruses, it has not been clear how ToLCNDV suddenly spread from the Indian subcontinent to the Mediterranean region. In 2017, ToLCNDV was diagnosed in young seedlings germinated naturally from fruits fallen in the prior year on a farm located in Giugliano in Campania, Naples, Italy, suggesting a possible role of the seeds in vertical transmission of the virus. Because sweet potato whiteflies were widespread naturally in that region, it was necessary to verify that in an artificial insect vector-free condition. Seeds were harvested from two ToLCNDV-infected zucchini squash cultivars in Naples in 2017 and 2018 to examine whether ToLCNDV can be transmitted from zucchini squash seeds to young plants. Viral DNA was amplified from these seeds and 1- to 3-week-old seedlings germinated from them with a ToLCNDV-specific primer set. According to PCR results, viral contamination was confirmed from all harvested seeds and dissemination was proven from 61.36% of tested seedling samples. Mechanical transmission from seed-borne virus-infected seedlings to healthy zucchini squash plants was also succesful, demonstrating that seedlings from ToLCNDV-infected seeds did act as inoculum. This is the first report demonstrating that ToLCNDV is a seed-transmissible virus in zucchini squash plants in Italy.
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http://dx.doi.org/10.3390/plants9050563DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7284533PMC
April 2020

The threat of seed-transmissible pepper yellow leaf curl Indonesia virus in chili pepper.

Microb Pathog 2020 Jun 10;143:104132. Epub 2020 Mar 10.

College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon, 16419, South Korea. Electronic address:

Recently, chili pepper (Capsicum annuum) plants in Indonesia have been devastated by a notorious bipartite begomovirus infection named Pepper yellow leaf curl Indonesia virus (PepYLCIV), which causes a distinct decrease in chili pepper production. Pepper yellow diseases have been known since early 2000; however, the spread of this virus thus far is distressing. These diseases can reduce chili yields by 20-100% in Indonesia. As previously known, begomovirus can be transmitted through whitefly to several host plants from the families Solanaceae, Compositae, and Leguminosae. In the field, a single plant was observed with severe symptoms of pepper yellow leaf curl disease, while other plants in the same field were asymptomatic and healthy. The observation leads to the possibility that the virus can be transmitted from previously infected chili pepper plants through seeds, as begomovirus transmission through seeds has been reported before. This study was conducted using seeds from chili peppers infected with viruses from different places in Indonesia. Whole seeds, embryos, and seedlings from PepYLCIV infected seeds were investigated in this study by performing viral genome DNA extraction, uracil DNA glycosylase-PCR, and sequencing analysis. Results revealed that both DNA-A and DNA-B of PepYLCIV in seeds and embryos of infected chili pepper plants were detected. The results also showed that 25-67% of PepYLCIV DNA-A and 50-100% of DNA-B were detected from seedlings grown from infected chili pepper seed collected from different location, thus confirming PepYLCIV as a seed-transmissible virus in chili pepper plants.
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http://dx.doi.org/10.1016/j.micpath.2020.104132DOI Listing
June 2020

Life Cycle-Based Host Range Analysis for Tomato Spotted Wilt Virus in Korea.

Plant Pathol J 2020 Feb 1;36(1):67-75. Epub 2020 Feb 1.

Highland Agriculture Research Institute, National Institute of Crop Science, Rural Development Administration, Pyeongchang 25342, Korea.

(TSWV) is one of the plant viruses transmitted by thrips and causes severe economic damage to various crops. From 2008 to 2011, to identify natural host species of TSWV in South Korea, weeds and crops were collected from 5 regions (Seosan, Yesan, Yeonggwang, Naju, and Suncheon) where TSWV occurred and were identified as 1,104 samples that belong to 144 species from 40 families. According to reverse transcription-polymerase chain reaction, TSWV was detected from 73 samples from 23 crop species, 5 of which belonged to family Solanaceae. Additionally, 42 weed species were confirmed as natural hosts of TSWV with three different life cycles, indicating that these weed species could play an important role as virus reservoirs during no cultivation periods of crops. This study provides up-to-date comprehensive information for TSWV natural hosts in South Korea.
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http://dx.doi.org/10.5423/PPJ.FT.12.2019.0290DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7012576PMC
February 2020

Identification of begomoviruses from different cryptic species of Bemisia tabaci in Bangladesh.

Microb Pathog 2020 Feb 13;142:104069. Epub 2020 Feb 13.

Division of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University, Daegu, Republic of Korea; Institute of Plant Medicine, Kyungpook National University, Daegu, Republic of Korea; Institute of Agricultural Science and Technology, Kyungpook National University, Daegu, Republic of Korea. Electronic address:

Bemisia tabaci is a global species complex consisting of at least 40 cryptic species. It is also a vector for at least 100 species of begomovirus, many of which cause severe crop damage. The relationship between begomoviruses and cryptic species of the B. tabaci species complex, however, remains unclear. Our previous study [13] was identified four cryptic species (Asia I, Asia II 1, Asia II 5, and Asia II 10) of B. tabaci from Bangladesh. Using those 110 whitefly samples, vector-based PCR analysis identified 8 different begomovirus species: BYVMV, BGYVV, OELCV, SLCCV, SLCV, TbCSV, ToLCBV, and ToLCNDV. The overall rate of virus infection was 26.4%, and BYVMV and ToLCNDV were the most frequently detected in the B. tabaci vector. Virus infection rates for Asia I, Asia II 1, Asia II 5, and Asia II 10 were 22.4% (15/67), 35% (7/20), 27.3% (6/22), and 100% (1/1), respectively. Each cryptic species infected multiple virus species, but SLCCV, TbCSV, and BGYVV were each only detected in, Asia I, Asia II 1, and Asia II 5, respectively. This study demonstrates the geographic distribution of various begomoviruses in Bangladesh and their relationships with cryptic species of B. tabaci.
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http://dx.doi.org/10.1016/j.micpath.2020.104069DOI Listing
February 2020

Enhanced Anticancer Effect of Adding Magnesium to Vitamin C Therapy: Inhibition of Hormetic Response by SVCT-2 Activation.

Transl Oncol 2020 Feb 31;13(2):401-409. Epub 2019 Dec 31.

Yeom Chang-Hwan Hospital, Seoul, 06605, Republic of Korea. Electronic address:

l-Ascorbic acid (vitamin C, AA) is known as an antioxidant, but at high concentrations, AA can kill cancer cells through a prooxidant property. Sodium-dependent vitamin C transporter family-2 (SVCT-2) determines the cellular uptake of AA, and the activity of SVCT-2 is directly related to the anticancer activity of AA. Cancer cells that showed high SVCT-2 expression levels were more sensitive to AA treatment than cancer cells with low SVCT-2 expression levels. Cells with low SVCT-2 expression showed a hormetic response to a low dose of AA. Magnesium ions, which are known to activate SVCT-2, could increase the V value of SVCT-2, so we investigated whether providing magnesium supplements to cancer cells with low SVCT-2 expression that had shown a hormetic response to AA would elevate the V value of SVCT-2, allowing more AA to accumulate. To evaluate the effects of magnesium on cancer cells, MgSO and MgCl were screened as magnesium supplements; both forms showed synergistic anticancer effects with AA. Taken together, the results of this study suggest that magnesium supplementation enhanced the anticancer effect of AA by inhibiting the hormetic response at a low dose. This study has also demonstrated that AA treatment with magnesium supplementation provided more effective anticancer therapy than AA treatment alone.
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http://dx.doi.org/10.1016/j.tranon.2019.10.017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6940627PMC
February 2020

Genome-wide identification of long non-coding RNAs in tomato plants irradiated by neutrons followed by infection with .

PeerJ 2019 28;7:e6286. Epub 2019 Jan 28.

Department of Physics, Sungkyunkwan University, Suwon, South Korea.

Long non-coding RNAs (lncRNAs) play an important role in regulating many biological processes. In this study, tomato seeds were first irradiated by neutrons. Eight tomato mutants were then selected and infected by (TYLCV). RNA sequencing followed by bioinformatics analyses identified 1,563 tomato lncRNAs. About half of the lncRNAs were derived from intergenic regions, whereas antisense lncRNAs accounted for 35%. There were fewer lncRNAs identified in our study than in other studies identifying tomato lncRNAs. Functional classification of 794 lncRNAs associated with tomato genes showed that many lncRNAs were associated with binding functions required for interactions with other molecules and localized in the cytosol and membrane. In addition, we identified 19 up-regulated and 11 down-regulated tomato lncRNAs by comparing TYLCV infected plants to non-infected plants using previously published data. Based on these results, the lncRNAs identified in this study provide important resources for characterization of tomato lncRNAs in response to TYLCV infection.
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http://dx.doi.org/10.7717/peerj.6286DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6354667PMC
January 2019

Transcriptome profiles of tomato plants after neutron irradiation and infection with TYLCV.

Physiol Plant 2019 Feb;165(2):427-441

Department of Physics, Sungkyunkwan University, Suwon 16419, Republic of Korea.

Ionizing radiation is ubiquitous in the environment and can cause mutagenesis in living organisms. In this study, we examined the effects of neutron irradiation on tomato plants. Neutron irradiation decreased tomato germination rates, but most irradiated tomato plants did not show any significant phenotype. However, tomato mutants infected by Tomato yellow leaf curl virus (TYLCV) displayed resistance against TYLCV compared to the wild type (WT), which showed disease symptoms. RNA-Seq data demonstrated that the expression profiles of eight tomato mutants were significantly different from that of the WT. The transcriptomes obtained from presoaked seeds were highly altered compared to those of dry seeds. Increased irradiation time resulted in severe changes in the tomato transcriptome; however, different neutron irradiation intensities affected the expressions of different sets of genes. A high number of single-nucleotide polymorphisms in tomato transcriptomes suggest that neutron irradiation strongly impacts plant transcriptomes. The transition/transversion values among mutants were almost constant and were lower than that of the non-irradiated sample (WT), suggesting that neutron irradiation caused an effect. Taken together, this is the first report showing the effects of neutron irradiation on tomato plants by transcriptome analyses.
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http://dx.doi.org/10.1111/ppl.12913DOI Listing
February 2019

Phylogeographic analysis of the full genome of Sweepovirus to trace virus dispersal and introduction to Korea.

PLoS One 2018 13;13(8):e0202174. Epub 2018 Aug 13.

Department of Genetic Engineering, Sungkyunkwan University, Suwon, Korea.

Sweet potato is a vegetatively propagated crop that is produced for both growth in Korean fields and for export out of the country. The viruses that are present in introduced sweet potatoes can spread both domestically and to foreign countries. Determining the time and path of virus movement could help curtail its spread and prevent future dispersal of related viruses. Determining the consequences of past virus and sweet potato dispersal could provide insight into the ecological and economic risks associated with other sweet potato-infecting viral invasions. We therefore applied Bayesian phylogeographic inferences and recombination analyses of the available Sweepovirus sequences (including 25 Korean Sweepovirus genomes) and reconstructed a plausible history of Sweepovirus diversification and movement across the globe. The Mediterranean basin and Central America were found to be the launchpad of global Sweepovirus dispersal. Currently, China and Brazil are acting as convergence regions for Sweepoviruses. Recently reported Korean Sweepovirus isolates were introduced from China in a recent phase and the regions around China and Brazil continue to act as centers of Sweepovirus diversity and sites of ongoing Sweepovirus evolution. The evidence indicates that the region is an epidemiological hotspot, which suggests that novel Sweepovirus variants might be found.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202174PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6089449PMC
February 2019

Hormetic dose response to -ascorbic acid as an anti-cancer drug in colorectal cancer cell lines according to SVCT-2 expression.

Sci Rep 2018 07 27;8(1):11372. Epub 2018 Jul 27.

Yeom Chang-Hwan hospital, Seoul, 06605, Republic of Korea.

-Ascorbic acid (vitamin C, AA) exhibits anti-cancer effects with high-dose treatment through the generation of reactive oxygen species (ROS) and selective damage to cancer cells. The anti-cancer effects of -ascorbic acid are determined by sodium-dependent vitamin C transporter 2 (SVCT-2), a transporter of -ascorbic acid. In this study, we demonstrate that -ascorbic acid treatment showed efficient anti-cancer activity in cell lines with high expression levels of SVCT-2 for a gradient concentration of -ascorbic acid from 10 μM -2 mM. However, in low SVCT-2 expressing cell lines, high-dose -ascorbic acid (>1 mM) showed anti-cancer effects but low-dose (<10 μM) treatment induced cell proliferation. Such conflicting results that depend on the concentration are called a hormetic dose response. A hormetic dose response to low-dose -ascorbic acid was also observed in high SVCT-2 expressing cell lines in the presence of a SVCT family inhibitor. Insufficient uptake of -ascorbic acid in low SVCT-2 expressing cancer cell lines cannot generate sufficient ROS to kill cancer cells, resulting in the hormetic response. Molecular analysis confirmed the increased expression of cancer proliferation markers in the hormetic dose response. These results suggest that -ascorbic exhibits a biphasic effect in cancer cells depending on SVCT-2 expression.
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http://dx.doi.org/10.1038/s41598-018-29386-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6063950PMC
July 2018

Phylogenetic Characterization of Population in Korea: Evidence of Reassortment between Isolates from Different Origins.

Plant Pathol J 2018 Jun 1;34(3):199-207. Epub 2018 Jun 1.

Crop Protection Division, National Academy of Agricultural Science, Rural Development Administration, Wanju 55365, Korea.

(ToCV) is a whitefly-transmitted and phloem-limited crinivirus. In 2013, severe interveinal chlorosis and bronzing on tomato leaves, known symptoms of ToCV infection, were observed in greenhouses in Korea. To identify ToCV infection in symptomatic tomato plants, RT-PCR with ToCV-specific primers was performed on leaf samples collected from 11 tomato cultivating areas where ToCV-like symptoms were observed in 2013 and 2014. About half of samples (45.18%) were confirmed as ToCV-infected, and the complete genome of 10 different isolates were characterized. This is the first report of ToCV occurring in Korea. The phylogenetic relationship and genetic variation among ToCV isolates from Korea and other countries were also analysed. When RNA1 and RNA2 are analysed separately, ToCV isolates were clustered into three groups in phylogenetic trees, and ToCV Korean isolates were confirmed to belong to two groups, which were geographically separated. These results suggested that Korean ToCV isolates originated from two independent origins. However, the RNA1 and RNA2 sequences of the Yeonggwang isolate were confirmed to belong to different groups, which indicated that ToCV RNA1 and RNA2 originated from two different origins and were reassorted in Yeonggwang, which is the intermediate point of two geographically separated groups.
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http://dx.doi.org/10.5423/PPJ.OA.10.2017.0220DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5985646PMC
June 2018

Probiotic Expressing a Nucleic Acid-Hydrolyzing Minibody (3D8 Scfv) Enhances Probiotic Activities in Mice Intestine as Revealed by Metagenomic Analyses.

Genes (Basel) 2018 May 29;9(6). Epub 2018 May 29.

Department of Genetic Engineering, Sungkyunkwan University, 2066, Seobu-ro, Jangan-gu, Suwon 16419, Korea.

Probiotics are well known for their beneficial effects for animals, including humans and livestock. Here, we tested the probiotic activity of expressing 3D8 scFv, a nucleic acid-hydrolyzing mini-antibody, in mice intestine. A total of 18 fecal samples derived from three different conditions at two different time points were subjected to high-throughput 16S ribosomal RNA (rRNA) metagenomic analyses. Bioinformatic analyses identified an average of 290 operational taxonomic units. After administration of , populations of the probiotics , , and increased, whereas the population of harmful bacteria such as species decreased. Furthermore, continuous administration of resulted in emerging as the dominant probiotic after competition with other existing probiotics. Expression of 3D8 scFv protein specifically increased the population of , which is another probiotic. In summary, our results showed that expressing 3D8 scFv protein enhanced probiotic activity in mice intestine with no observable side effects. Thus, the system developed in this study may be a good tool for the expression of recombinant protein using probiotics.
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http://dx.doi.org/10.3390/genes9060276DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6027128PMC
May 2018

Seed Transmission of in White Soybean ().

Plant Pathol J 2017 Aug 1;33(4):424-428. Epub 2017 Aug 1.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 16419, Korea.

(TYLCV) infection of the common bean () has been reported, but soybean () has not previously been identified as a TYLCV host. Five cultivars of white soybean were agro-inoculated using an infectious TYLCV clone. At 30 days post-inoculation, they showed infection rates of 25% to 100%. Typical TYLCV symptoms were not observed in any inoculated plants. To examine whether TYLCV was transmitted in soybean seeds, DNA was isolated from bundles of five randomly selected seeds from TYLCV-inoculated soybean plants and amplified with a TYLCV-specific primer set. With the exception of one bundle, all bundles of seeds were verified to be TYLCV-infected. Virus dissemination was also confirmed in three of the 14 bunches. Viral replication was also identified in seeds and seedlings. This is the first report demonstrating that soybean is a TYLCV host, and that TYLCV is a seed-transmissible virus in white soybean.
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http://dx.doi.org/10.5423/PPJ.NT.02.2017.0043DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5538446PMC
August 2017

Hydrolysis of Hyaluronic Acid in Lymphedematous Tissue Alleviates Fibrogenesis via T1 Cell-Mediated Cytokine Expression.

Sci Rep 2017 02 24;7(1):35. Epub 2017 Feb 24.

Department of Genetic Engineering, Sungkyunkwan University, Suwon, 16419, Republic of Korea.

Although surgery and radiation are beneficial for treating cancer, they can also lead to malfunctions of the lymphatic system such as secondary lymphedema. This abnormality of the lymphatic system is characterized by severe swelling, adipogenesis, inflammation, and fibrosis in the lymphedematous region. Moreover, the proliferation of fibrotic tissue in the lymphedematous region generates edema that is no longer spontaneously reversible. No treatment for fibrosis has been validated in patients with lymphedema. In our efforts to develop a therapeutic agent for lymphedema fibrosis, we used a newly established mouse hind limb model. Previous studies have demonstrated that hyaluronic acid accumulates in the lymphedematous region. Thus, we challenged mice with of hyaluronidase (HYAL), with the aim of reducing fibrogenesis. After subcutaneous injections in the lymphedematous mouse leg every two days, the volume of lymphedema had reduced significantly by 7 days post-operation. Histochemical analysis indicated that collagen accumulation and myofibroblast differentiation were decreased in epidermal tissues after HYAL injection. Moreover, it was associated with upregulation of interferon-gamma, increased numbers of Th1 cells, and downregulation of interleukin-4 and interleukin-6 in the lymphedematous region and spleen. These results indicate that hydrolysis of hyaluronic acid can boost an anti-fibrotic immune response in the mouse lymphedema model.
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http://dx.doi.org/10.1038/s41598-017-00085-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5428353PMC
February 2017

Tomato yellow leaf curl virus (TYLCV-IL): a seed-transmissible geminivirus in tomatoes.

Sci Rep 2016 Jan 8;6:19013. Epub 2016 Jan 8.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.

Tomato yellow leaf curl virus (TYLCV) is one of the most well-known tomato-infecting begomoviruses and transmitted by Bemisia tabaci. Seed transmission has previously been reported for some RNA viruses, but TYLCV has not previously been described as a seed-borne virus. In 2013 and 2014, without whitefly-mediated transmission, TYLCV was detected in young tomato plants germinated from fallen fruits produced from TYLCV-infected tomato plants in the previous cultivation season. In addition, TYLCV-Israel (TYLCV-IL) was also detected in seeds and their seedlings of TYLCV-infected tomato plants that were infected by both viruliferous whitefly-mediated transmission and agro-inoculation. The seed infectivity was 20-100%, respectively, and the average transmission rate to seedlings was also 84.62% and 80.77%, respectively. TYLCV-tolerant tomatoes also produced TYLCV-infected seeds, but the amount of viral genome was less than seen in TYLCV-susceptible tomato plants. When tomato plants germinated from TYLCV-infected seeds, non-viruliferous whiteflies and healthy tomato plants were placed in an insect cage together, TYLCV was detected from whiteflies as well as receiver tomato plants six weeks later. Taken together, TYLCV-IL can be transmitted via seeds, and tomato plants germinated from TYLCV-infected seeds can be an inoculum source of TYLCV. This is the first report about TYLCV seed transmission in tomato.
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http://dx.doi.org/10.1038/srep19013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4705557PMC
January 2016

Preventive Activity against Influenza (H1N1) Virus by Intranasally Delivered RNA-Hydrolyzing Antibody in Respiratory Epithelial Cells of Mice.

Viruses 2015 Sep 21;7(9):5133-44. Epub 2015 Sep 21.

Department of Genetic Engineering, Sungkyunkwan University, 2066, Seobu-ro, Jangan-gu, Suwon 16419, Korea.

The antiviral effect of a catalytic RNA-hydrolyzing antibody, 3D8 scFv, for intranasal administration against avian influenza virus (H1N1) was described. The recombinant 3D8 scFv protein prevented BALB/c mice against H1N1 influenza virus infection by degradation of the viral RNA genome through its intrinsic RNA-hydrolyzing activity. Intranasal administration of 3D8 scFv (50 μg/day) for five days prior to infection demonstrated an antiviral activity (70% survival) against H1N1 infection. The antiviral ability of 3D8 scFv to penetrate into epithelial cells from bronchial cavity via the respiratory mucosal layer was confirmed by immunohistochemistry, qRT-PCR, and histopathological examination. The antiviral activity of 3D8 scFv against H1N1 virus infection was not due to host immune cytokines or chemokines, but rather to direct antiviral RNA-hydrolyzing activity of 3D8 scFv against the viral RNA genome. Taken together, our results suggest that the RNase activity of 3D8 scFv, coupled with its ability to penetrate epithelial cells through the respiratory mucosal layer, directly prevents H1N1 virus infection in a mouse model system.
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http://dx.doi.org/10.3390/v7092863DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4584307PMC
September 2015

Detection of the dinoflagellate, Cochlodinium polykrikoides, that forms algal blooms using sandwich hybridization integrated with nuclease protection assay.

Biotechnol Lett 2016 Jan 3;38(1):57-63. Epub 2015 Sep 3.

South Sea Environment Research Department, Korea Institute of Ocean Science & Technology, Geoje, 656-830, Korea.

Objectives: To detect Cochlodinum polykrikoides in long-term monitoring and high-throughput sampling projects using an integrated sandwich hybridization and nuclease protection assay (NPA-SH).

Results: The specificity of the probes was verified with individual and mixed cultures as well as field collection, and the quantity of C. polykrikoides determined by NPA-SH analysis showed a good correlation with that determined by cell-counting with a light microscope. In addition a standard curve for C. polykrikoides was established to represent the correlation between optical absorbance in the NPA-SH assay and cell density.

Conclusions: This approach provides an efficient alternative to traditional, morphology-based methods for the rapid identification and quantification of harmful algal species and could be used to monitor phytoplankton in field surveys.
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http://dx.doi.org/10.1007/s10529-015-1947-5DOI Listing
January 2016

Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity.

Mol Cells 2015 Sep 7;38(9):773-80. Epub 2015 Aug 7.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Korea.

3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 LD50 PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% (5 μg) and 47% (10 μg). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.
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http://dx.doi.org/10.14348/molcells.2015.0073DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588720PMC
September 2015

Seasonal Dynamics of Marine Microbial Community in the South Sea of Korea.

PLoS One 2015 29;10(6):e0131633. Epub 2015 Jun 29.

South Sea Environment Research Department, Korea Institute of Ocean Science and Technology, Geoje, 656-830, Republic of Korea; Korea University of Science and Technology, Daejeon, 305-350, Republic of Korea.

High-resolution 16S rRNA tag pyrosequencing was used to obtain seasonal snapshots of the bacterial diversity and community structure at two locations in Gosung Bay (South Sea, Korea) over a one year period. Seasonal sampling from the water column at each site revealed highly diverse bacterial communities containing up to 900 estimated Operational Taxonomic Units (OTUs). The Alphaproteobacteria and Gammaproteobacteria were the most abundant groups, and the most frequently recorded OTUs were members of Pelagibacter and Glaciecola. In particular, it was observed that Arcobacter, a genus of the Epsilonproteobacteria, dominated during summer. In addition, Psedoalteromonadaceae, Vibrionaceae and SAR11-1 were predominant members of the OTUs found in all sampling seasons. Environmental factors significantly influenced the bacterial community structure among season, with the phosphate and nitrate concentrations contributing strongly to the spatial distribution of the Alphaproteobacteria; the Gammaproteobacteria, Flavobacteria, and Actinobacteria all showed marked negative correlations with all measured nutrients, particularly silicon dioxide and chlorophyll-a. The results suggest that seasonal changes in environmental variables contribute to the dynamic structure of the bacterial community in the study area.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0131633PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4487691PMC
April 2016

Oxaliplatin-Induced Peripheral Neuropathy via TRPA1 Stimulation in Mice Dorsal Root Ganglion Is Correlated with Aluminum Accumulation.

PLoS One 2015 30;10(4):e0124875. Epub 2015 Apr 30.

Department of Genetic Engineering, Sungkyunkwan University, Suwon, Korea.

Oxaliplatin is a platinum-based anticancer drug used to treat metastatic colorectal, breast, and lung cancers. While oxaliplatin kills cancer cells effectively, it exhibits several side effects of varying severity. Neuropathic pain is commonly experienced during treatment with oxaliplatin. Patients describe symptoms of paresthesias or dysesthesias that are triggered by cold (acute neuropathy), or as abnormal sensory or motor function (chronic neuropathy). In particular, we found that aluminum levels were relatively high in some cancer patients suffering from neuropathic pain based on clinical observations. Based on these findings, we hypothesized that aluminum accumulation in the dorsal root ganglion (DRG) in the course of oxaliplatin treatment exacerbates neuropathic pain. In mice injected with oxaliplatin (three cycles of 3 mg/kg i.p. daily for 5 days, followed by 5 days of rest), we detected cold allodynia using the acetone test, but not heat hyperalgesia using a hot plate. However, co-treatment with aluminum chloride (AlCl3∙6H2O; 7 mg/kg i.p. for 14 days: equivalent 0.78 mg/kg of elemental Al) and oxaliplatin (1 cycle of 3 mg/kg i.p. daily for 5 days, followed by 5 days of rest) synergistically induced cold allodynia as well as increased TRPAl mRNA and protein expression. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis showed a significant increase in aluminum concentrations in the DRG of mice treated with aluminum chloride and oxaliplatin compared to aluminum chloride alone. Similarly, in a mouse induced-tumor model, aluminum concentrations were increased in DRG tissue and tumor cells after oxaliplatin treatment. Taken together, these findings suggest that aluminum accumulation in the DRG may exacerbate neuropathic pain in oxaliplatin-treated mice.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0124875PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4415919PMC
December 2015

Tomato yellow leaf curl virus Can Overwinter in Stellaria aquatica, a Winter-Hardy TYLCV-Reservoir Weed.

Plant Dis 2015 May 30;99(5):588-592. Epub 2015 Apr 30.

Department of Genetic Engineering, Sungkyunkwan University.

Tomato yellow leaf curl virus (TYLCV), one of the most serious plant viruses in tropical and subtropical regions, is transmitted to host plants by the vector insect Bemisia tabaci. In order to control TYLCV, it is important to identify weed hosts for overwintering TYLCV. Stellaria aquatica, a winter-hardy weed, was found growing with TYLCV-infected tomato plants in greenhouse production. TYLCV was detected in S. aquatica plants by polymerase chain reaction and Southern blot hybridization analysis. The intergenic region nucleotide sequences amplified from TYLCV-infected tomato plants, TYLCV-viruliferous whiteflies, and S. aquatica were identical. During winter (December to February), TYLCV-viruliferous whiteflies and TYLCV-infected tomato plants were removed or absent from greenhouses. However, S. aquatica plants were observed over a period of 10 months from August to May in such greenhouses, and TYLCV was consistently detected in some of these plants. To investigate the transmission of TYLCV from TYLCV-infected S. aquatica plants to healthy tomato plants by whiteflies, TYLCV-infected S. aquatica plants were transplanted to pots in cages with nonviruliferous whiteflies and healthy tomato plants. After 4 weeks, tomato plants developed typical TYLCV disease symptoms, and TYLCV was detected in both whiteflies and tomato plants. These results show that S. aquatica can act as a winter-hardy reservoir for TYLCV, and suggest that this weed could play an important role in overwintering of TYLCV in tomato greenhouses.
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http://dx.doi.org/10.1094/PDIS-04-14-0352-REDOI Listing
May 2015

Fatty acid methyl ester profiles and nutritive values of 20 marine microalgae in Korea.

Asian Pac J Trop Med 2015 Mar;8(3):191-6

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.

Objective: To screen the fatty acid (FA) composition of 20 marine microalgae species, including seven Diophyceae, six Bacillariophyceae, four Chlorophyceae, two Haptophyceae and one Raphidophyceae species.

Methods: Microalgal cells cultured at the Korea Institute of Ocean Science & Technology were harvested during the late exponential growth phase and the FA composition analyzed.

Results: The FA composition of microalgae was species-specific. For example, seven different species of Dinophyceae were composed primarily of C14:0, C16:0, C18:0, C20:4n-6, C20:5n-3 and C22:6n-3, while C14:0, C16:0, C16:1, C18:0, C20:5n-3 and C22:6n-3 were abundant FAs in six species of Bacillariophyceae. In addition, four Chlorophyceae, two Haptophyceae and one Raphidophyceae species all contained a high degree of C16:1n-7 [(9.28-34.91)% and (34.48-35.04)%], C14:0 [(13.34-25.96)%] and [(26.69-28.24)%], and C16:0 [(5.89-29.15)%] and [(5.70-16.81)%]. Several factors contribute to the nutritional value of microalgae, including the polyunsaturated FA content and n-3 to n-6 FA ratio, which could be used to assess the nutritional quality of microalgae.

Conclusions: This study is the first comprehensive assessment of the FA composition and nutritional value of microalgae species in South Korea, and identifies the potential utility of FAs as species-specific biomarkers.
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http://dx.doi.org/10.1016/S1995-7645(14)60313-8DOI Listing
March 2015

Advanced loop-mediated isothermal amplification method for sensitive and specific detection of Tomato chlorosis virus using a uracil DNA glycosylase to control carry-over contamination.

J Virol Methods 2015 Mar 4;213:68-74. Epub 2014 Dec 4.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address:

In 2013, Tomato chlorosis virus (ToCV) was identified in symptomatic tomato plants in Korea. In the present study, a loop-mediated isothermal amplification (LAMP) method was developed using four specific primers designed against ORF6 in ToCV RNA2 to detect ToCV rapidly and with high sensitivity. The optimized reaction involved incubation of a reaction mixture containing 2U Bst DNA polymerase and 4mM MgSO4 for 1h at 60-62 °C. Although specific and rapid detection of ToCV by LAMP was confirmed, false-positive reactions caused by carry-over contamination sometimes occurred because of the high sensitivity of LAMP compared with other detection methods. To prevent false-positive reactions, dUTP was substituted for dTTP and uracil-DNA glycosylase (UDG) was added to the LAMP reaction. First, the LAMP reaction was conducted successfully with substitution of dUTP for dTTP. Before the next reaction, LAMP products with incorporated dUTP were cleaved selectively by UDG without any effect on thymine-containing DNA (template DNA). This modified LAMP method complemented with UDG treatment to prevent carry-over contamination offers a potentially powerful method for detecting plant viruses.
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http://dx.doi.org/10.1016/j.jviromet.2014.10.020DOI Listing
March 2015

Sweet pepper confirmed as a reservoir host for tomato yellow leaf curl virus by both agro-inoculation and whitefly-mediated inoculation.

Arch Virol 2014 Sep 30;159(9):2387-95. Epub 2014 Apr 30.

Department of Genetic Engineering, Sungkyunkwan University, 300 Cheoncheon-dong, Jangan-gu, Suwon, 440-746, Republic of Korea.

Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, has a single-stranded DNA genome. TYLCV can induce severe disease symptoms on tomato plants, but other hosts plants such as cucurbits and peppers are asymptomatic. A full-length DNA clone of a Korean TYLCV isolate was constructed by rolling-circle amplification from TYLCV-infected tomatoes in Korea. To assess relative susceptibility of sweet pepper varieties to TYLCV, 19 cultivars were inoculated with cloned TYLCV by agro-inoculation. All TYLCV-infected sweet peppers were asymptomatic, even though Southern hybridization and polymerase chain reaction analysis showed TYLCV genomic DNA accumulation in roots, stems, and newly produced shoots. Southern hybridization indicated that TYLCV replicated and moved systemically from agro-inoculated apical shoot tips to roots or newly produced shoots of sweet peppers. Whitefly-mediated inoculation experiments showed that TYLCV can be transmitted to tomatoes from TYLCV-infected sweet peppers. Taken together, these results indicate that sweet pepper can be a reservoir for TYLCV in nature.
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http://dx.doi.org/10.1007/s00705-014-2072-9DOI Listing
September 2014

Lamium amplexicaule (Lamiaceae): a weed reservoir for tomato yellow leaf curl virus (TYLCV) in Korea.

Arch Virol 2014 Jun 11;159(6):1305-11. Epub 2013 Dec 11.

Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea.

After the first identification of tomato yellow leaf curl virus (TYLCV) in the southern part of Korea in 2008, TYLCV has rapidly spread to tomato farms in most regions of Korea. From 2008 to 2010, a survey of natural weed hosts that could be reservoirs of TYLCV was performed in major tomato production areas of Korea. About 530 samples were collected and identified as belonging to 25 species from 11 families. PCR and Southern hybridization were used to detect TYLCV in samples, and replicating forms of TYLCV DNA were detected in three species (Achyranthes bidentata, Lamium amplexicaule, and Veronica persica) by Southern hybridization. TYLCV transmission mediated by Bemisia tabaci from TYLCV-infected tomato plants to L. amplexicaule was confirmed, and TYLCV-infected L. amplexicaule showed symptoms such as yellowing, stunting, and leaf curling. TYLCV from infected L. amplexicaule was also transmitted to healthy tomato and L. amplexicaule plants by B. tabaci. The rate of infection of L. amplexicaule by TYLCV was similar to that of tomato. This report is the first to show that L. amplexicaule is a reservoir weed host for TYLCV.
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http://dx.doi.org/10.1007/s00705-013-1913-2DOI Listing
June 2014

Loop-mediated isothermal amplification for the rapid detection of Chrysanthemum chlorotic mottle viroid (CChMVd).

J Virol Methods 2013 Oct 4;193(1):232-7. Epub 2013 Jun 4.

Department of Genetic Engineering, Sungkyunkwan University, Suwon 440-746, Republic of Korea.

Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method offering rapid, sensitive, and convenient diagnosis of infectious diseases. Chrysanthemum chlorotic mottle viroid (CChMVd) causes one of the most serious viral diseases in chrysanthemum in Korea. A sensitive LAMP assay was developed for rapidly detecting CChMVd infection. The assay was based on a set of four primers matching the specific region of the CChMVd genome. The CChMVd LAMP primer sets were designed using the sequences from nonsymptomatic and symptomatic CChMVd isolates in Korea. The efficiency and specificity of this method were optimized using Bst DNA polymerase, which allowed for increased viroid detection sensitivity. The reaction was carried out at 65 °C for 90 min, and was improved by adding SYBR Green I dye to the inside of the reaction tube lid prior to amplification. The results indicate that this LAMP method will be useful for chrysanthemum viroid disease monitoring and detecting CChMVd infectious disease.
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http://dx.doi.org/10.1016/j.jviromet.2013.05.016DOI Listing
October 2013
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