• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.290-290
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• 2022

Powdery mildew is known to be one of the serious diseases in C. moschata cultivation. Plants infected with powdery mildew cause damage to cultivation areas such as occurrence of deformity fruit and decrease in quantity. also, it has been reported that many farms have difficulties in controlling powdery mildew due to the outbreak under various conditions throughout the year. Therefore, this study intends to perform a phenotype test and a genotype test for C. moschata 60 lines grown in Jenong S&T. Podospareaxanthii, known as a pathogen that causes powder mildew disease in pumpkins in Korea, was collected and used as an inoculation source, phenotype test was performed by examining the infection area rate(%) of powdery mildew disease that occurred in leaves 25 days after inoculation. It was determined that 0% of the infection area rate was in the first stage, 1 to 5% in the second stage, 6 to 15% in the third stage, 16 to 30% in the fourth stage, and 31% or more in the fifth stage, The first and second stages were judged as resistance, the third as moderate resistance, and the fourth and fifth stages as sensitivity. As a result of the phenotype test, it was confirmed that the resistance was 21 points, moderate resistance was 14 points, and sensitivity was 25 points. After searching for the genes related to powdery mildew resistance resistance, pm-0, CmbHLH87, and LOC111453072, 21 points of resistance and 9 points of moderate resistance identified through phenotype tests were identified through gel electrophoresis after polymerase chain reaction(PCR) using 5 primers related to 3 genes. As a result of genotype testing of a total 30 points, the CmbHLH87 and LOC111453072 gene were found to be resistant bands in all points, PMR1 was identified as 20 points for resistance, 4 points for moderate resistance, and 6 points for sensitivity, PMR2 was not identified in the entire band, and PMR5 was identified as 18 point for resistance, 3 points for moderate resistance, and 9 points for sensitivity. As a result, when comparing the phenotype test results and genotype test results, CmbHLH87 and LOC111453072 genes was 100% consistent in resistance and moderate resistance, PMR1 was 95.2% in resistance, 44.4% in moderate resistance, and PMR5 was 90% in resistance and 33.3% in moderate resistance, PMR2 was not consistent in resistance and moderate resistance. Therefore, it is expected that more accurate PMR test will be possible by using molecular markers(PMR1, PMR5) and by developing CmbHLH87 and LOC111453072 gene-related molecular markers.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.14-21
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• 2011

Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.251-256
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• 2005

Cytokinins are essential plant hormones that play crucial roles in various aspects of plant growth and development. To better understand the molecular mechanisms of cytokinin action, we identified cytokinin related proteins by a proteomic approach. Proteins extracted from control and trans-zeatin treated Arabidopsis seedlings were separated and analyzed by two dimensional gel analysis. Differentially expressed protein spots were identified with peptide mass fingerprinting based on matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and database searching, We obtained ten up-regulated and one down-regulated proteins upon t-zeatin treatment. The expression of the following proteins was induced; pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase and IAA amino acid hydrolase homolog.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7085-7090
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• 2013

Objective: Published data have shown that microRNAs (miRNAs) could play a potential role as diagnostic and prognostic indicators in cancers. Data for the predictive value of microRNA-155 are inconclusive. The aim of the present analysis was therefore to evaluate the role of miR-155 in prognosis for patients with a variety of carcinomas. Methods: Relevant studies were identified by searching PubMed and EMBASE. Data were extracted from studies comparing overall survival (OS), recurrence-free survival (RFS) or cancer-specific survival (CSS) in patients with carcinoma with higher miR-155 expression and those with lower levels. The pooled hazard ratios (HRs) and 95% confidence intervals (CIs) of miR-155 for clinical outcome were calculated. Results: A total of 15 studies were included. The pooled hazard ratio (HR) for OS of higher miR-155 expression in cancerous tissue was 1.89 (95% CI: 1.20-2.99, P=0.006), which could markedly predict poorer survival in general cancer. For RFS/CSS, elevated miR-155 was also associated with poor prognosis of cancer (HR=1.50, 95% CI: 1.10-2.05, P=0.01). On subgroup analysis, the pooled HR for OS in non-small cell lung cancer (NSCLC) was 2.09 (95% CI: 0.68-6.41, P > 0.05), but for RFS/CSS was 1.28 (95% CI: 1.05-1.55, P=0.015), with statistical significance; the pooled HRs for OS and RFS/CSS in digestive system neoplasms were 3.04 (95% CI: 1.48-6.24, P=0.003) and 2.61 (95% CI: 1.98-3.42, P<0.05), respectively. Conclusions: The results indicated that the miR-155 expression level plays a prognostic role in patients with cancer, especially NSCLCs and digestive system carcinomas.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.14-14
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• 2010

Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

• Journal of KIISE:Databases
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• v.34 no.2
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• pp.119-132
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• 2007

In molecular biology, approximate subsequence search is one of the most important operations. In this paper, we propose an accurate and efficient method for approximate subsequence search in large DNA databases. The proposed method basically adopts a binary trie as its primary structure and stores all the window subsequences extracted from a DNA sequence. For approximate subsequence search, it traverses the binary trie in a breadth-first fashion and retrieves all the matched subsequences from the traversed path within the trie by a dynamic programming technique. However, the proposed method stores only window subsequences of the pre-determined length, and thus suffers from large post-processing time in case of long query sequences. To overcome this problem, we divide a query sequence into shorter pieces, perform searching for those subsequences, and then merge their results. To verify the superiority of the proposed method, we conducted performance evaluation via a series of experiments. The results reveal that the proposed method, which requires smaller storage space, achieves 4 to 17 times improvement in performance over the suffix tree based method. Even when the length of a query sequence is large, our method is more than an order of magnitude faster than the suffix tree based method and the Smith-Waterman algorithm.

• Journal of Animal Science and Technology
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• v.45 no.5
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• pp.731-738
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• 2003

The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

• KSBB Journal
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• v.24 no.3
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• pp.312-320
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• 2009

Stable cell preservation is an essential factor in the regenerative medicine for cell therapies and transplantation of biologic materials. In this study, we studied to provide more stable hypothermic preservation by protection of cell damage during the preservation at $4^{\circ}C$. The result of searching for key components that have excellent efficacy in hypothermic preservation of cells, we have identified the fact that the hypothermic preservation adding protein hydrolysates such as yeast hydrolysate is far superior to others. All protein hydrolysates that are derived from animal, plant and microbe sources have superior efficacy, especially the peptides which have molecular weights under 10 kDa have the best efficacy among the components of protein hydrolysate. The protein hydrolysates prevented the decrease of ATP level in the cells caused by hypothermic environment and they inhibited the generation of ROS. Adding antioxidants and control agents of osmotic pressure were showed to have more superior efficacy in hypothermic preservation. Finally, KUL261 solution (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%), the preservation solution developed in this study, showed the best efficacy in both cell viability and cell growth more than other conventional preservation solutions. In conclusion, the improved hypothermic preservation solution that contains the protein hydrolysates as a key component provide the best preservation efficacy. It provides better efficacy than other preservation solutions and will contribute to both the development of regenerative medicine and global commercialization in this therapeutic field.

• Applied Biological Chemistry
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• v.38 no.3
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• pp.212-217
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• 1995

This study was aimed for determining the biochemical mechanism of cold tolerance in crops and for searching the biochemical genetic marker related with cold tolerance by the analysis of isozyme pattern. We investigated various biochemical changes induced by the long-term cold acclimation in cold sensitive rape (B. napus) and in cold tolerant 'Sandongchae'(B. campestris) seedlings. The cold shock after long-term cold acclimation to B. napus and B. campestris greatly increased the activities of peroxidase 157% and 50% in root fraction and, 201% and 205% in hypocotyl, respectively. Simultaneously, the activity of superoxide dismutase was largely increased in hypocotyl fraction, too. Protein contents of hypocotyl fractions in B. napus and B. campestris were also increased by 11.4% and 57.8%, respectively. The band of pl 6.4 among peroxidase isozymes newly biosynthesized during long-term cold acclimation was emerged in the hypocotyl fraction of cold tolerant B. campestris as well as in the root of both species. From above and previous results, we presented a model of interconversions of molecular oxygen species due to the cold injury and biochemically inferred the mechanism of cold tolerance in crops.

• Horticultural Science & Technology
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• v.33 no.5
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• pp.722-729
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• 2015

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.