• The Plant Pathology Journal
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• v.37 no.2
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• pp.99-114
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• 2021

Tan spot of wheat, caused by Pyrenophora tritici-repentis (Ptr), results in a yield loss through chlorosis and necrosis of healthy leaf tissue. The major objective of this study was to compare gene expression in resistant and susceptible wheat cultivars after infection with Ptr ToxA-producing race 2 and direct infiltration with Ptr ToxA proteins. Greenhouse experiments included exposure of the wheat cultivars to pathogen inoculum or direct infiltration of leaf tissue with Ptr-ToxA protein isolate. Samples from the experiments were subjected to RNA sequencing. Results showed that ToxA RNA sequences were first detected in samples collected eight hours after treatments indicating that upon Ptr contact with wheat tissue, Ptr started expressing ToxA. The resistant wheat cultivar, in response to Ptr inoculum, expressed genes associated with plant resistance responses that were not expressed in the susceptible cultivar; genes of interest included five chitinases, eight transporters, five pathogen-detecting receptors, and multiple classes of signaling factors. Resistant and susceptible wheat cultivars therefore differed in their response in the expression of genes that encode chitinases, transporters, wall-associated kinases, permeases, and wound-induced proteins, among others. Plants exposed to Ptr inoculum expressed transcription factors, kinases, receptors, and peroxidases, which are not expressed as highly in the control samples or samples infiltrated with ToxA. Several of the differentially expressed genes between cultivars were found in the Ptr resistance QTLs on chromosomes 1A, 2D, 3B, and 5A. Future studies should elucidate the specific roles these genes play in the wheat response to Ptr.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.62 no.1
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• pp.40-50
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• 2017

Seed storage proteins are used as carbon and nitrogen sources for the nutritional improvement of seeds. Since the composition of proteins from the Korean cultivars of proso millet is unknown, this study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from proso millet seeds of various cultivars were investigated using proteomic techniques such as 2-D electrophoresis coupled with mass fingerprinting; 1152 (differentially expressed) protein spots were detected on the 2-D gels. Among them, 26 reproducible protein spots were analyzed using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Out of the 26 proteins, 2 proteins were upregulated in all the millet cultivars, while 13 proteins were upregulated and 11 proteins were downregulated in 2 cultivars. Abundance of most of the identified protein species associated with polysaccharide and starch metabolism, transcription, and pathogenesis was significantly enhanced, while that of other protein species involved in glycolysis, stress response, and transduction was severely reduced. Taken together, the results suggest that the differential expression of the proteins from the four millet cultivars may be cultivar-specific. By conducting a proteomic investigation of millet seeds from different cultivars, we sought to better understand the functional categorization of individual proteins on the basis of their molecular functions. We believe that the identified proteins may help in investigating genetic variations in millet cultivars.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.120-120
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• 2017

Since the composition of proteins from the Korean cultivars of Proso millet is unknown; thereby, the present study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from the millet seeds of various cultivars, were investigated using proteomic techniques as 2D electrophoresis coupled with mass fingerprinting. The 1152 (differentially expressed) proteins were detected on 2-D gel. Among them, 26 reproducible protein spots were analyzed by MALDI-TOF-TOF mass spectrometry. Out of 26 proteins, 2 proteins were up-regulated towards all cultivars of millet, while 7 proteins were up-regulated and 13 proteins were down-regulated against only one cultivar. However, abundance in most identified protein species, associated with metabolism, transcription and transcription was significantly enhanced, while that of another protein species involved in polysaccharide metabolism, stress response and pathogenesis were severely reduced. Taken together, the results observed from the study suggest that the differential expression of proteins from the four cultivars of millet may be cultivar-specific. Taken together, a proteomic investigation of millet seeds from different cultivars, we sought to better understand the genetic variation of millet cultivars representing the future millet research, and the functional categorization of individual proteins on the basis of their molecular function.

• Journal of fish pathology
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• v.22 no.3
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• pp.383-389
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• 2009

Analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics studies. As part of studies on the immune system of olive flounder, a total of 251 EST sequences from gill cDNA library were generated to identify and characterize important genes in the immune machanisms of olive flounder. Of the 251 clones, 126 clones (50.2%) were identified as orthologues of known genes from olive flounder and other organisms. Among the 126 EST clones, 16 clones (12.7%) were representing 9 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 clones (79.4%) representing 103unique genes were identified as orthologs of known genes from other organisms. We also identified several kinds of immune associated proteins, indicating EST as a powerful method for identifying immune related genes of fish as well as identifying novel genes. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.181-187
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• 2013

In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.128-128
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• 2017

Copper (Cu) is very toxic to plant cells due to its inhibitory effects on many physiological and biochemical processes. In spite of its potential physiological and economic significance, molecular characterization after Cu stress has so far been grossly overlooked in sorghum. To explore the molecular alterations that occur in response to copper stress, the present study was executed in ten-day-old Cu-exposed leaves of sorghum seedlings. The growth of shoots was markedly reduced, and ionic alterations were prominently observed in the leaves when the seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CuSO_4$. Using two-dimensional gels with silver staining, 643 differentially expressed protein spots (${\geq}1.5-fold$) were identified as either significantly increased or reduced in abundance. Of these spots, a total of 24 protein spots (${\geq}1.5-fold$) from Cu-exposed sorghum leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Of the 24 differentially expressed proteins from Cu-exposed sorghum leaves, a total of 13 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of most identified protein species, which function in carbohydrate metabolism, stress defense, and protein translation, was significantly enhanced, while that of another protein species involved in energy metabolism, photosynthesis and growth and development were severely reduced. The resulting differences in protein expression patterns together with related morpho-physiological processes suggested that these results could help to elucidate plant adaptation to Cu stress and provide insights into the molecular mechanisms of Cu responses in $C_4$ plants. The over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.125-125
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• 2017

The roots of Platycodon grandiflorum are commonly used for treating bronchitis, asthma, tuberculosis, diabetes, and other inflammatory diseases. Since the molecular mechanism underlying the roots of the plant is unclear. Therefore, the present study was conducted to profile proteins from liquid cultured tetraploid roots of Platycodon grandi orum fl using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 659 differentially expressed proteins were identified from the liquid medium cultured tetraploid roots of which 32 proteins spots (${\geq}1.5-fold$) were sorted for mass spectrometry analysis. Out of these 32 proteins, a total of 15 proteins were up-regulated such as Serine carboxypeptidase-like 27, Transcription factor bHLH150, 60 kDa jasmonate-induced protein, Cytosolic Fe-S cluster assembly factor NBP35, Regulatory associated protein of TOR 2 and a total of 17 proteins were down-regulated such as Protein G1-like2, Phenylalanine ammonia-lyase, Fructokinase-2, Trihelix transcription factor GT-3a, Guanine nucleotide-binding protein alpha-1 subunit. However, the frequency distribution of identified proteins was carried out within functional categories based on molecular functions, cellular components, and biological processes. Functional categorization revealed that the most of the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase, transferase activity, protein binding and hydrolase activity. In addition, the proteomic feedback of tetraploid roots of P. grandiflorum may potentially be used to understand the characteristics of proteins and their functions.

• Molecules and Cells
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• v.21 no.3
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• pp.381-388
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• 2006

Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

• Mycobiology
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• v.38 no.4
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• pp.302-309
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• 2010

Formic acid is a representative carboxylic acid that inhibits bacterial cell growth, and thus it is generally considered to constitute an obstacle to the reuse of renewable biomass. In this study, Saccharomyces cerevisiae was used to elucidate changes in protein levels in response to formic acid. Fifty-seven differentially expressed proteins in response to formic acid toxicity in S. cerevisiae were identified by 1D-PAGE and nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) analyses. Among the 28 proteins increased in expression, four were involved in the MAP kinase signal transduction pathway and one in the oxidative stress-induced pathway. A dramatic increase was observed in the number of ion transporters related to maintenance of acid-base balance. Regarding the 29 proteins decreased in expression, they were found to participate in transcription during cell division. Heat shock protein 70, glutathione reductase, and cytochrome c oxidase were measured by LC-MS/MS analysis. Taken together, the inhibitory action of formic acid on S. cerevisiae cells might disrupt the acidbase balance across the cell membrane and generate oxidative stress, leading to repressed cell division and death. S. cerevisiae also induced expression of ion transporters, which may be required to maintain the acid-base balance when yeast cells are exposed to high concentrations of formic acid in growth medium.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.113-113
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• 2003

A number of studies have demonstrated recently nonthermal interactions of extremely low frequency electromagnetic fields with cellular systems, such as the cells of the immune system. Particular concern came from epidemiological findings, which correlated environmental exposure of human body to weak electromagnetic fields with an elevated risk for developing certain type of leukemias and cancers. Several home/environmental sources generating extremely low frequency electromagnetic fields, such as 50 - 60 Hz high-voltage transmission lines, video display terminals, electric blankets, clinical nuclear magnetic resonance imaging procedures, etc., may interact with the human body. In this study we examined the effects of static magnetic fields (SMF) on the phagocytosis of the murine peritoneal macrophages (C57BL/6). The cells were exposed in vitro for 24 h at 37$^{\circ}C$ to 400 G SMF. The phagocytic activity of peritoneal macrophages was determined with a luminometer. Exposure to the SMF decreased phagocytic activity of murine peritoneal macrophages. In order to provide a more exact mechanism of the phenomenon, we analyzed peritoneal macrophages for alteration in their proteomes. The expression levels of these 5 proteins were increased in the SMF. In total 5 proteins which were differentially expressed in the SMF compared with control group were identified. The expression levels of these 5 proteins were increased in the SMF.