• Title/Summary/Keyword: in silico

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Identification of Potential DREB2C Targets in Arabidopsis thaliana Plants Overexpressing DREB2C Using Proteomic Analysis

  • Lee, Kyunghee;Han, Ki Soo;Kwon, Young Sang;Lee, Jung Han;Kim, Sun Ho;Chung, Woo Sik;Kim, Yujung;Chun, Sung-Sik;Kim, Hee Kyu;Bae, Dong-Won
    • Molecules and Cells
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    • v.28 no.4
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    • pp.383-388
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    • 2009
  • The dehydration responsive element binding protein 2C (DREB2C) is a dehydration responsive element/C-repeat (DRE/CRT)-motif binding transcription factor that induced by mild heat stress. Previous experiments established that overexpression of DREB2C cDNA driven by the cauliflower mosaic virus 35S promoter (35S:DREB2C) resulted in increased heat tolerance in Arabidopsis. We first analyzed the proteomic profiles in wild-type and 35S:DREB2C plants at a normal temperature ($22^{\circ}C$), but could not detect any differences between the proteomes of wild-type and 35S: DREB2C plants. The transcript level of DREB2C in 35S: DREB2C plants after treatment with mild heat stress was increased more than two times compared with expression in 35S:DREB2C plants under unstressed condition. A proteomic approach was used to decipher the molecular mechanisms underlying thermotolerance in 35S:DREB2C Arabidopsis plants. Eleven protein spots were identified as being differentially regulated in 35S:DREB2C plants. Moreover, in silico motif analysis showed that peptidyl-prolyl isomerase ROC4, glutathione transferase 8, pyridoxal biosynthesis protein PDX1, and elongation factor Tu contained one or more DRE/CRT motifs. To our knowledge, this study is the first to identify possible targets of DREB2C transcription factors at the protein level. The proteomic results were in agreement with transcriptional data.

Screening of Genetic Polymorphisms of CYP3A4 and CYP3A5 Genes

  • Lee, Jin Sol;Cheong, Hyun Sub;Kim, Lyoung Hyo;Kim, Ji On;Seo, Doo Won;Kim, Young Hoon;Chung, Myeon Woo;Han, Soon Young;Shin, Hyoung Doo
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.6
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    • pp.479-484
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    • 2013
  • Given the CYP3A4 and CYP3A5's impact on the efficacy of drugs, the genetic backgrounds of individuals and populations are regarded as an important factor to be considered in the prescription of personalized medicine. However, genetic studies with Korean population are relatively scarce compared to those with other populations. In this study, we aimed to identify CYP3A4/5 polymorphisms and compare the genotype distributions among five ethnicities. To identify CYP3A4/5 SNPs, we first performed direct sequencing with 288 DNA samples which consisted of 96 Koreans, 48 European-Americans, 48 African-Americans, 48 Han Chinese, and 48 Japanese. The direct sequencing identified 15 novel SNPs, as well as 42 known polymorphisms. We defined the genotype distributions, and compared the allele frequencies among five ethnicities. The results showed that minor allele frequencies of Korean population were similar with those of the Japanese and Han Chinese populations, whereas there were distinct differences from European-Americans or African-Americans. Among the pharmacogenetic markers, frequencies of $CYP3A4^*1B$ (rs2740574) and $CYP3A5^*3C$ (rs776742) in Asian groups were different from those in other populations. In addition, minor allele frequency of $CYP3A4^*18$ (rs28371759) was the highest in Korean population. Additional in silico analysis predicted that two novel non-synonymous SNPs in CYP3A5 (+27256C>T, P389S and +31546T>G, I488S) could alter protein structure. The frequency distributions of the identified polymorphisms in the present study may contribute to the expansion of pharmacogenetic knowledge.

Brain Wave Control Effect of Smart-wave via Docking into the Odorant-binding Protein (스마트 웨이브 조성물질의 odorant 결합 단백질에 대한 분자 결합 친화도 비교 분석 및 후각 흡입으로 유도되는 뇌파 변화 연구)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.26 no.3
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    • pp.346-352
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    • 2016
  • Aroma inhalation therapy has traditionally been used not only in alternative medicinal treatment but also in psychotherapy. In the first stage of the study, the in silico molecular binding affinity of the major ingredients of Smart-Wave (SW) on the active site of the odorant-binding protein (OBP) was compared with that of citrate anions. The binding affinity of the chemical mixture formula of the major ingredients of SW on the OBP was relatively higher than that of citrate anions. In addition, nasal inhalation of SW had a positive effect upon changes in brain waves. Eighteen healthy volunteers participated in the experiment. The study consisted of measurements of the brain’s meditation level recordings in the pre- and post-SW inhalation periods as compared with negative (EV) and positive (HB) control groups. After SW inhalation, all the subjects stated that they felt “fresher” and that the SW trial group had significantly changed the brain’s meditation in a positive way. SW inhalation also converted EV-induced unstable brain meditation wave patterns into more stable patterns. Collectively, the results of this empirical study strongly suggest that the SW mixture activates the OBP and controls the mental state by regulating brain waves. The results provide scientific evidence that the SW formula has potential as an effective mental-stress controller.

In-silico annotation of the chemical composition of Tibetan tea and its mechanism on antioxidant and lipid-lowering in mice

  • Ning Wang ;Linman Li ;Puyu Zhang;Muhammad Aamer Mehmood ;Chaohua Lan;Tian Gan ;Zaixin Li ;Zhi Zhang ;Kewei Xu ;Shan Mo ;Gang Xia ;Tao Wu ;Hui Zhu
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.682-697
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    • 2023
  • BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice. MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH2O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected. Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS: This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

Identification and in silico analysis of two types of serpin genes from expressed sequence tags (ESTs) of the Oriental land snail, Nesiohelix samarangae (동양달팽이 (Nesiohelix samarangae) 의 expressed sequence tags (ESTs) 로부터 분리한 2종류의 Serpin 유전자 분석)

  • Park, So Young;Jeong, Ji Eun;Hwang, Hee Ju;Wang, Tae Hun;Park, Eun Bi;Kim, Yong Min;Lee, Jun-Sang;Han, Yeon Soo;Yang, Seung-Ha;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.30 no.2
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    • pp.155-163
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    • 2014
  • Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.

Microbial Forensics: Comparison of MLVA Results According to NGS Methods, and Forensic DNA Analysis Using MLVA (미생물법의학: 차세대염기서열분석 방법에 따른 MLVA 결과 비교 및 이를 활용한 DNA 감식)

  • Hyeongseok Yun;Seungho Lee;Seunghyun Lim;Daesang Lee;Sehun Gu;Jungeun Kim;Juhwan Jeong;Seongjoo Kim;Gyeunghaeng Hur;Donghyun Song
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.4
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    • pp.507-515
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    • 2024
  • Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

Bioinformatic Analysis of NLS (Nuclear Localization Signals)-containing Proteins from Mollusks (생물정보학을 이용한 연체동물의 NLS (Nuclear Localization Signals) 포함 단백질의 분석)

  • Lee, Yong-Seok;Kang, Se-Won;Jo, Yong-Hun;Gwak, Heui-Chul;Chae, Sung-Hwa;Choi, Sang-Haeng;Ahn, In-Young;Park, Hong-Seog;Han, Yeon-Soo;Kho, Weon-Gyu
    • The Korean Journal of Malacology
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    • v.22 no.2
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    • pp.109-113
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    • 2006
  • Subcellular localization of a protein containing nuclear localization signals (NLS) has been well studied in many organisms ranging from invertebrates to vertebrates. However, no systematic analysis of NLS-containing proteins available from Mollusks has been reported. Here, we describe in silico screening of NLS-containing proteins using the mollusks database that contains 22,138 amino acids. To screen putative proteins with NLS-motif, we used both predict NLS and perl script. As a result, we have found 266 proteins containing NLS sequences which are about 1.2% out of the entire proteins. On the basis of KOG (The eukaryotic orthologous groups) analysis, we can't predict the precise functions of the NLS-containing proteins. However, we found out that these proteins belong to several types of proteins such as chromatin structure and dynamics, translation, ribosomal structure, biogenesis, and signal transduction mechanism. In addition, we have analysed these sequences based on the classes of mollusks. We could not find many from the species that are the main subjects of phylogenetic studies. In contrast, we noticed that cephalopods has the highest number of NLS-containing proteins. Thus, we have constructed mollusks NLS database and added these information and data to the mollusks database by constructing web interface. Taken together, these information will be very useful for those who are or will be studying NLS-containing proteins from mollusks.

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Study on Mo(V) Species, Location and Adsorbates Interactions in MoH-SAPO-34 by Employing ESR and Electron Spin-Echo Modulation Spectroscopies (ESR, ESEM을 이용한 이온 교환된 MoH-SAPO-34에 대한 Mo의 화학종, 위치 및 흡착상호작용에 관한 연구)

  • Back, Gern-Ho;Jang, Chang-Ki;Ru, Chang-Kuk;Cho, Young-Hwan;So, Hyun-Soo;Kevan, Larry
    • Journal of the Korean Chemical Society
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    • v.46 no.1
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    • pp.26-36
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    • 2002
  • A solid-state reaction of $MoO_3$ with as-synthesized H-SAPO-34 generated paramagnetic Mo(V) species. The dehydration resulted in weak Mo(V) species, and subsequent activation resulted in the formation of Mo(V) species such as $Mo(V)_{5c}$ and $Mo(V)_{6c}$ that are characterized by ESR. The data of ESR and ESEM show the oxomolybdenum species, to be $(MoO_2)^+$ or $(MoO)^{3+}$. The $(MoO_2)^+$ species seems to be more probable. Since H-SAPO-34 has a low framework negative charge, $(MoO)^{3+}$ with a high positive charge can not be easily stabilized. A solution reaction between the solution of silico-molybdic acid and calcined H-SAPO-34 resulted in only $(MoO_2)^+$ species. A rhombic ESR signal is observed on adsorption of $D_2O$, $CD_3OH$, $CH_3Ch_2OD$ and $ND_3$. The Location and coordination structure of Mo(V) species has been determined by three-pulse electron spin-echo modulation data and their simulations. After the adsorption of methanol, ethylene, ammonia, and water for MoH-SAPO-34, three molecules, one molecule, one and one molecule, respectively, are directly coordinated to $(MoO_2)^+)$.

Sequence Analysis of a Cryptic Plasmid pKW2124 from Weissella cibaria KLC140 and Construction of a Surface Display Vector

  • Kim, Soo Young;Oh, Chang Geun;Lee, Young Joo;Choi, Kyu Ha;Shin, Doo Sik;Lee, Si Kyung;Park, Kab Joo;Shin, Hakdong;Park, Myeong Soo;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.545-554
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    • 2013
  • Plasmid isolation of kimchi-derived Weissella cibaria KLC140 revealed six different plasmids. The smallest plasmid, pKW2124, was DNA sequenced and characterized, showing 2,126 bp with a GC content of 36.39% and five putative open reading frames (ORFs). In silico analysis of these ORFs showed ORF1 encodes a putative replication protein similar to rolling circular replication proteins from other lactic acid bacteria. However, a single-stranded intermediate was not detected when S1 nuclease was treated, suggesting it may follow theta replication. Interestingly, the replication initiation site of this plasmid is 100% identical to other plasmids from lactic acid bacteria, suggesting it may function for replication initiation. To construct a surface layer expression vector, pTSLGFP, slpA encoding the surface layer protein from Lactobacillus acidophilus was PCR amplified and fused with the gfp gene, forming a SLGFP fused gene. The plasmid pKW2124 was cloned into the XbaI site of pUC19, forming an Weissella-E. coli shuttle vector pKUW22. NheI-linearized pTSLGFP was ligated into pKUWCAT containing pKUW22 and the chloramphenicol acetyltransferase gene from pEK104, resulting in an 8.6 kb pKWCSLGFP surface layer expression vector. After transformation of this vector into W. cibaria KLC140, a GFP fluorescence signal was detected on the surface of the transformant, substantiating production of SLGFP fused protein and its secretion. This is the first report for construction of a Weissella surface layer expression vector, which may be useful for surface layer production of beneficial proteins in Weissella.

Rough Computational Annotation and Hierarchical Conserved Area Viewing Tool for Genomes Using Multiple Relation Graph. (다중 관계 그래프를 이용한 유전체 보존영역의 계층적 시각화와 개략적 전사 annotation 도구)

  • Lee, Do-Hoon
    • Journal of Life Science
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    • v.18 no.4
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    • pp.565-571
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    • 2008
  • Due to rapid development of bioinformatics technologies, various biological data have been produced in silico. So now days complicated and large scale biodata are used to accomplish requirement of researcher. Developing visualization and annotation tool using them is still hot issues although those have been studied for a decade. However, diversity and various requirements of users make us hard to develop general purpose tool. In this paper, I propose a novel system, Genome Viewer and Annotation tool (GenoVA), to annotate and visualize among genomes using known information and multiple relation graph. There are several multiple alignment tools but they lose conserved area for complexity of its constrains. The GenoVA extracts all associated information between all pair genomes by extending pairwise alignment. High frequency conserved area and high BLAST score make a block node of relation graph. To represent multiple relation graph, the system connects among associated block nodes. Also the system shows the known information, COG, gene and hierarchical path of block node. In this case, the system can annotates missed area and unknown gene by navigating the special block node's clustering. I experimented ten bacteria genomes for extracting the feature to visualize and annotate among them. GenoVA also supports simple and rough computational annotation of new genome.