• Journal of Marine Life Science
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• v.3 no.2
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• pp.51-58
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• 2018

Variations in water temperature are known to affect almost every part of fish physiology. The rise in water temperature due to climate change can physically damage fish. This study was conducted to evaluate the health status of the Atlantic salmon (Salmo salar) at high water temperature (20℃) than the optimum water temperature (15℃). Liver tissue exerts important metabolic functions in thermal adaptation. Therefore, liver tissue was used in this study. The evaluation method is to develop the biomarker gene using NGS RNAseq analysis and to examine the expression pattern using RT-qPCR analysis. The NGS RNAseq analysis revealed 1,366 differentially expressed genes, among which 880 genes were increase expressed and 486 genes were decrease expressed. The biomarker genes are such as heat shock protein 90 alpha (Hsp90α), heat shock protein 90 beta (Hsp90β) and cytochrome P450 1A (CYP1A). The selected genes are sensitive to changes in water temperature through NGS RNAseq analysis. Expression patterns of these genes through RT-qPCR were similar to those of NGS RNAseq analysis. The results of this study can be applied to other fish species and it is considered to be useful industrially.

• Journal of Internet Computing and Services
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• v.18 no.5
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• pp.47-53
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• 2017

In past few years, high-throughput sequencing, big-data generation, cloud computing, and computational biology are revolutionary. RNA sequencing is emerging as an attractive alternative to DNA microarrays. And the methods for constructing Gene Regulatory Network (GRN) from RNA-Seq are extremely lacking and urgently required. Because GRN has obtained substantial observation from genomics and bioinformatics, an elementary requirement of the GRN has been to maximize distinguishable genes. Despite of RNA sequencing techniques to generate a big amount of data, there are few computational methods to exploit the huge amount of the big data. Therefore, we have suggested a novel gene selection algorithm combining Support Vector Machines and Intensity-dependent normalization, which uses log differential expression ratio in RNAseq. It is an extended variation of support vector machine recursive feature elimination (SVM-RFE) algorithm. This algorithm accomplishes minimum relevancy with subsets of Big-Data, such as NCBI-GEO. The proposed algorithm was compared to the existing one which uses gene expression profiling DNA microarrays. It finds that the proposed algorithm have provided as convenient and quick method than previous because it uses all functions in R package and have more improvement with regard to the classification accuracy based on gene ontology and time consuming in terms of Big-Data. The comparison was performed based on the number of genes selected in RNAseq Big-Data.

• Korean Journal of Environmental Biology
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• v.36 no.2
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• pp.131-139
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• 2018

The warming of water as a result of climate change affects fish habitat. Variations in water temperature affect fish physiology almost totally. The rise in water temperature due to climate change leads to hypoxia following decreased oxygen solubility and decreased binding capacity of oxygen-carrying hemoglobin. This study was conducted to evaluate the health status of Atlantic salmon (Salmo salar) fry at elevated water temperatures($20^{\circ}C$) compared with optimum water temperature ($15^{\circ}C$). The method facilitated the detection of biomarker genes using NGS RNAseq analysis and evaluation of their expression pattern using RT-qPCR analysis. The biomarker genes included interferon alpha-inducible protein 27-like protein 2A transcript variant X3, protein L-Myc-1b-like, placenta growth factor-like transcript variant X1, fibroblast growth factor receptor-like 1 transcript variant X1, transferrin, intelectin, thioredoxin-like, c-type lectin lectoxin-Thr1-like, ladderlectin-like and calponin-1. The selected biomarker genes were sensitive to changes in water temperature based on NGS RNAseq analysis. The expression patterns of these genes based on RT-qPCR were similar to those of NGS RNAseq analysis.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.249-259
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• 2013

Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::${\Omega}$) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::${\Omega}$). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.

• Proceedings of the Korean Society of Crop Science Conference
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• 2020.11a
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• pp.107-107
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• 2020

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.10a
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• pp.217-217
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• 2017

• Plant Breeding and Biotechnology
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• v.6 no.4
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• pp.413-425
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• 2018

A previous work developed and identified a new omega-5 gliadin deficient wheat line named O-free by crossing Keumkang and Olgeuru, which is nutritionally quite meaningful in that omega-5 gliadin is one of the known wheat allergens. To verify the characteristics of the O-free, we performed RNA sequencing (RNAseq) analysis of the O-free and the two parent lines (Keumkang and Olgeuru). The results of the similarity analysis with the ESTs for gliadins and glutenins showed that the O-free ESTs had no similarity with the omega-5 gliadin sequences but had similarity to other gliadins and glutenins. Furthermore, mapping results between the raw RNAseq data from the O-free and the omega-5 gliadin sequence showed a clear deletion of the N-terminal sequences which are an important signature of omega-5 gliadin. We also designed specific PCR primers that could identify omega-5 gliadin in the genomic DNA. The results showed that no omega-5 gliadin fragments were detected in the O-free. According to these results, we confirmed that the deficiency of omega-5 gliadin in the O-free is not caused by post-transcriptional or post-translational regulations such as epigenetic phenomena but by a simple deletion in the chromosome. Furthermore, we showed that the low-molecular weight glutenin subunit (LMW-GS) gene in the O-free had a single nucleotide polymorphism (SNP) causing a premature stop codon, resulting in a truncated polypeptide. We expect that the O-free line may serve as an excellent source of wheat that could prevail in the hypo-allergen wheat market, which has recently gained interest world-wide.

• Molecules and Cells
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• v.46 no.10
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• pp.611-626
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• 2023

Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.117-117
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• 2017

The onion (Allium cepa L.) is the most widely cultivated species of the genus Allium, especially it has been valued because of the pungent flavor and aroma. Allium species including onion has very large genome sizes ranging from approximately 10 to 20 Gbp, which have complicated genomic studies and precluded genome sequencing until recently. A population of 186 F2 individuals derived from a cross of 'Umjinara' ${\times}$ 'Sinsunhwang' and the two parental lines were used for this study. For the development of framework map, various types of markers including SSRs, RAPD, SNPs, and CAPS makers have been used for polymorphism test. Especially, a lot of SNP and CAPS loci were developed from the onion transcriptome sequence by RNASEQ of two parental lines. The GBS libraries have been constructed based on a modified protocol from Poland Lab using a two-enzyme system. We have been developing markers showing polymorphism between two parental lines, and genotyping for all F2 individuals were finished for a number of polymorphic markers. For the construction of GBS libraries, a set of 192 barcoded adapters were generated from complementary oligonucleotides with XhoI overhang sequence and unique barcodes of length 4-8 bp and they have been tested using two parental linesto determine the optimum conditions for GBS analysis.

• Proceedings of the Korean Society of Crop Science Conference
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• 2019.09a
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• pp.148-148
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• 2019

Wheat is a very important crop as a food source worldwide, but gluten in wheat causes a variety of allergic reactions. Previous studies have developed ${\omega}-5$ gliadin deleted O-free, known as the central antigen of WDEIA (wheat-dependent exercise-induced anaphylaxis). In this study, we performed RNA sequencing on the grains and leaves of the allergic-reduced species O-free and their cultivars, Keumkang and Olgeuru, to analyze differentially expressed genes (DEG) based on different cultivars and tissues. Tissues of all species were biologically repeated three times. We used bowtie2 version 2.3.5.1 to get sequence data from RNAseq and used cufflinks and Tophat programs to find DEG. When comparing leaf and grain tissues, a total of 1,244 DEGs were found in the leaf tissues while only 563 DEGs were found in the grain tissues. As a result of gene ontology analysis of differentially expressed genes, the leaf tissues were mostly included in the "catalytic activity" part of molecular function, "metabolic process" part of biological process, and "membrane" part of cell component. The grain tissues were mostly included in the "metabolic process" part of biological process, "binding" and "catalytic activity" part of molecular function, and "membrane, cell, cell part" parts of cell component. Based on these results, we present information on the differentially expressed genes of the three cultivars of leaves and grains. This study could be an important basis for studying the characteriztion of O-free.