• The Plant Pathology Journal
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• v.32 no.3
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• pp.173-181
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• 2016

Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1), which is associated with fumonisin B1 bio-synthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi.

• Bulletin of the Korean Chemical Society
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• v.35 no.7
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• pp.2197-2200
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• 2014

• Journal of Plant Biotechnology
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• v.43 no.3
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• pp.293-301
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• 2016

High-quality gene sets are necessary for functional research of genes. Although Perilla is a commonly cultivated oil crop and vegetable crop in Southeast Asia, the quality of its available gene set is insufficient. To construct a high-quality Perilla gene set, we sequenced mRNAs extracted from different tissues of Perilla citriodora, the wild species (2n = 20) of Perilla. To make a high-quality gene set for P. citriodora, we compared the quality of assemblies produced by Velvet and Trinity, the two well-known de novo assemblers, and improved the de novo assembly pipeline by optimizing k-mers and removing redundant sequences. We then selected representative transcripts for loci according to several criteria. The improved assembly yielded a total of 86,396 transcripts and 38,413 representative transcripts. We evaluated the assembled transcripts by comparing them to 638 homologous Arabidopsis genes involved in fatty acid and TAG biosynthesis pathways. High proportions of full-length genes and transcripts in the assembled transcripts matched known genes in other species, indicating that the P. citriodora gene set can be applied in future functional studies. Our study provides a reference P. citriodora gene set for further studies. It will serve as valuable genetic resource to elucidate the molecular basis of various metabolisms.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.68-71
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• 1999

To illustrate the action mechanism of pencycuron on Rhizoctonia solani, two experiments were conducted including the comparison of amino acids of $\beta$-tubulin between R-C (sensitive isolate) and Rh-131 (non-sensitive isolate), and the inhibitory effect of pencycuron on tubulin assembly in vitro. Both $\beta$-tubulin genes of R-C and Rh-131 proved to have 1,582 nucleotides encoding a protein of 445 amino acids, showing 98% homology in amino acid sequences between them. It was found that codons at 103, 236, and 267 for lysine (AGG), valine (GTC) and isoleucine (ATT) in R-C were replaced by codons for methionine (ATG), isoleucine (ATT) and methionine (ATG) in Rh-131, respectively. No inhibitory effect of pencycuron on the tubulin assembly was observed. It suggests that pencycuron may have no direct inhibitory effects on the assembly of tubulin at least in vitro.

• 한국균학회소식:학술대회논문집
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• 2018.05a
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• pp.22-22
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• 2018

Fungal genome sequencing and assembly have been trivial in these days. Genome analysis relies on high quality of gene prediction and annotation. Automatic fungal genome annotation pipeline is essential for handling genomic sequence data accumulated exponentially. However, building an automatic annotation procedure for fungal genomes is not an easy task. FunGAP (Fungal Genome Annotation Pipeline) is developed for precise and accurate prediction of gene models from any fungal genome assembly. To make high-quality gene models, this pipeline employs multiple gene prediction programs encompassing ab initio, evidence-, and homology-based evaluation. FunGAP aims to evaluate all predicted genes by filtering gene models. To make a successful filtering guide for removal of false-positive genes, we used a scoring function that seeks for a consensus by estimating each gene model based on homology to the known proteins or domains. FunGAP is freely available for non-commercial users at the GitHub site (https://github.com/CompSynBioLab-KoreaUniv/FunGAP).

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.194-194
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• 2022

Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.200-200
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• 2022

Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

• Journal of the Korea Society of Computer and Information
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• v.25 no.8
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• pp.1-8
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• 2020

In this paper, we propose a gene family based RNA-seq read distribution method in means to accelerate the overal transcriptome assembly computation time. To measure the performance of our transcriptome sequence data distribution method, we evaluated the performance by testing four types of data sets of the Arabidopsis thaliana genome (Whole Unclassified Reads, Family-Classified Reads, Model-Classified Reads, and Randomly Classified Reads). As a result of de novo transcript assembly in distributed nodes using model classification data, the generated gene contigs matched 95% compared to the contig generated by WUR, and the execution time was reduced by 4.2 times compared to a single node environment using the same resources.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.9
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• pp.1331-1341
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• 2001

Dairy cows are prone to fatty liver during the time of periparturient. Despite of the extensive studies, etiology and solutions for fatty liver are still not well known.The liver synthesizes triglycerides (TG) using precursors from bloodstream and secretes TG in form of very low density lipoprotein (VLDL) into bloodstream for the utilization by peripheral tissues. When the amount of TG synthesis exceeds the amount of secretion in VLDL-TG, TG accumulation within the liver occurs. Hepatic VLDL assembly and secretion involve multi-biochemical events.The availabilities of apolipoprotein B (apoB), E (apoE), microsomal triglyceride transfer protein (MTP) and soluble low density lipoprotein (LDL) receptor are now believed to be some of the main regulators for hepatic VLDL assembly and secretion. Studies in transgenic animals show that overexpression of these proteins stimulates VLDL production and secretion, which provides a possibility for alleviating bovine fatty liver by gene therapy. However, many problems remain to be solved to attain this goal. This review focuses on the molecular mechanisms of hepatic VLDL assembly and secretion, and the possibilities and problems of applying the knowledges to solve bovine fatty liver by gene therapy.

• BMB Reports
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• v.46 no.6
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• pp.310-315
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• 2013

The gene order on the X chromosome of eutherians is generally highly conserved, although an increase in the rate of rearrangement has been reported in the rodent lineage. Conservation of the X chromosome is thought to be caused by selection related to maintenance of dosage compensation. However, we herein reveal that the cattle (Btau4.0) lineage has experienced a strong increase in the rate of X-chromosome rearrangement, much stronger than that previously reported for rodents. We also show that this increase is not matched by a similar increase on the autosomes and cannot be explained by assembly errors. Furthermore, we compared the difference in two cattle genome assemblies: Btau4.0 and Btau6.0 (Bos taurus UMD3.1). The results showed a discrepancy between Btau4.0 and Btau6.0 cattle assembly version data, and we believe that Btau6.0 cattle assembly version data are not more reliable than Btau4.0.