• Title/Summary/Keyword: in silico

Search Result 387, Processing Time 0.026 seconds

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
    • /
    • v.23 no.4
    • /
    • pp.545-554
    • /
    • 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
    • /
    • v.18 no.4
    • /
    • pp.565-571
    • /
    • 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.

The Study on Analysis Method and Performance Evaluation of Portable GC/MS (휴대용 GC/MS 성능검사 및 분석방법 연구)

  • Park, Yuonshin;Noh, Hyeran;Yang, Heeseon;Seok, Gwanseol
    • Journal of the Society of Disaster Information
    • /
    • v.9 no.3
    • /
    • pp.249-258
    • /
    • 2013
  • Recieved Mar. 06, 2013 Revised Mar. 29, 2013 Accepted Sep. 11, 2013In this study, we evaluated the method performance and the optimum sample injection method of the portable GC/MS(HAPSITE) which were designed to operate on scene of chemical accidents. Chemicals used for performance test were vinyl chloride(VC), methyl chloride(MC), benzene(Bz), toluene(Tol). CRM(Certified Reference Material) Manufactured by KRISS(Korea Research Institute of Standards and Science) was used as the standard gas. The results showed that 1) Among three sample injection methods(Cylinder, Silico canister, and Bag), bag was identified as the most appropriate sample injection method. Bag material and capacity did not significantly affect the results. The most ideal reproducibility occurred at the 2-minute point of the purge time in the loop and tenax methods. 2) Performance evaluation of HAPSITE was conducted. Evaluation items were reproducibility, linearity, method detection limit, etc. Tenax concentration method was appropriate for larger molecules and Carbopack concentration method was suitable for smaller molecules.

Tyrosinase Inhibition-mediated Anti-melanogenic Effects by Catechin Derivatives Extracted from Ulmus parvifolia (참느릅나무에서 추출된 catechin 유도체 화합물의 멜라닌 생성 억제 효과)

  • Taehyeok Hwang;Hyo Jung Lee;Dong-Min Kang;Kyoung Mi Moon;Jae Cheal Yoo;Mi-Jeong Ahn;Dong Kyu Moon;Dong Kyun Woo
    • Journal of Life Science
    • /
    • v.33 no.2
    • /
    • pp.169-175
    • /
    • 2023
  • As a protective defensive mechanism against ultraviolet (UV) light exposure in skin tissue, melanocytes produce the pigment melanin. Tyrosinase plays a key role in melanin production in melanocytes. However, the overproduction of melanin can lead to lesions, such as freckles and dark spots. Thus, it is clinically important to find a modulating molecule to control melanogenesis by regulating tyrosinase expression and/or activity. It is known that catechin, a plant flavonoid, can reduce melano- genesis through the downregulation of tyrosinase expression. Here, we tested whether catechin derivatives isolated from the stem bark of Ulmus parvifolia have an effect on melanin production by regulating tyrosinase in mouse melanoma cells and in vitro mushroom tyrosinase. The catechin derivatives used in this study included C5A, C7A, C7G, and C7X. Treatments using these catechin derivatives reduced melanin production in mouse melanoma B16F10 cells in which melanogenesis was stimulated by α-MSH. Notably, the anti-melanogenic effects of catechin derivatives were similar to those of kojic acid, a well-known anti-melanogenic molecule. Both C5A and C7A directly inhibited the activity of tyrosinase isolated from mushrooms in vitro. Furthermore, our in silico computational simulation showed that these two compounds were expected to bind to the active site of tyrosinase, which is similar to kojic acid. In addition, all four catechin derivatives reduced tyrosinase protein expression. In summary, our results showed that catechin derivatives can reduce melanogenesis by regulating tyrosinase activity or expression. Thus, this study suggests that catechin derivatives isolated from U. parvifolia can be novel modulators of melanin production.

Anti-inflammatory Activities of Antimicrobial Peptide Locustacin Derived from Locusta migratoria in LPS-stimulated RAW264.7 Cells (풀무치 유래 항균 펩타이드 locustacin의 항염증 활성)

  • Choi, Ra-Yeong;Lee, Joon Ha;Seo, Minchul;Kim, In-Woo;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
    • /
    • v.31 no.10
    • /
    • pp.898-904
    • /
    • 2021
  • Locusta migratoria is a widespread locust species in many parts of the world and is considered an alternative source for the production of protein for value-added ingredients. We previously identified putative antimicrobial peptides derived from L. migratoria through an in silico analysis of its transcriptome. However, its anti-inflammatory effect has not been studied. In this study, we investigated the anti-inflammatory activities of the antimicrobial peptide locustacin (KTHILSFFPSFLPLFLKK-NH2) derived from L. migratoria on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Locustacin (50, 100, and 200 ㎍/ml) significantly reduced the production of nitric oxide (NO) in LPS-stimulated macrophages without any cytotoxicity. Locustacin also inhibited the mRNA and protein expression of pro-inflammatory mediators, such as inducible NO synthase and cyclooxygenase-2, in contrast to the presence of LPS alone. Locustacin decreased the release of LPS-induced pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, and their gene expression in a dose-dependent manner. Furthermore, locustacin (100 and/or 200 ㎍/ml) inhibited phosphorylation levels of extracellular signal regulated kinase, p38, and c-Jun N-terminal kinase. Locustacin also suppressed the degradation of inhibitory kappa B alpha, which was considered to be an inhibitor of nuclear factor kappa B (NF-κB). Collectively, these results demonstrate that locustacin can exert anti-inflammatory effects through the inhibition of mitogen-activated protein kinase (MAPK) phosphorylation, activation of NF-κB, and downstream inflammatory mediators in LPS-stimulated macrophage cells.

Variation in bioactive principles and bioactive compounds of Rosa rugosa fruit during ripening (해당화 열매 성숙단계에 따른 생리활성 및 기능성 물질 변화 분석)

  • Kwak, Minjeong;Eom, Seung Hee;Gil, Jinsu;Kim, Ju-Sung;Hyun, Tae Kyung
    • Journal of Plant Biotechnology
    • /
    • v.46 no.3
    • /
    • pp.236-245
    • /
    • 2019
  • Fruit ripening is a genetically programmed process involving a number of biochemical and physiological processes assisted by variations in gene expression and enzyme activities. This process generally affects the phytochemical profile and the bioactive principles in fruits and vegetables. To appraise the variation in bioactive principles of fruits from Rosa rugosa during its ripening process, we analyzed the changes in antioxidant and anti-elastase activities and polyphenolic compounds during the four ripening stages of fruits. Overall, an extract of unripe fruits contained the highest levels of total phenolic and flavonoid contents, radical scavenging activity, reducing power, oxygen radical antioxidant capacity, and elastase inhibitory activity, compared with the extracts of fruits at other stages of ripening. Additionally, we found that the reduction of flavonoid content occurs because of decreased transcriptional levels of genes involved in flavonoid biosynthesis pathway during the ripening process. Based on HPLC analysis, we found that the extract of unripe fruits contained the highest amount of myricetin, caffeic acid, chlorogenic acid, syringic acid, and p-coumaric acid and suggested that the antioxidant and anti-elastase activities of the extract obtained from stage 1, should be mediated by the presence of these compounds. Additionally, we analyzed the interaction sites and patterns between these compounds and elastase using the structure-based molecular docking approach, and suggested that chlorogenic acid strongly interacted with elastase. Together, these findings suggest that the maturity of fruits has profound effects on the pharmaceutical value of R. rugosa.

Bioinformatic Analysis of the Canine Genes Related to Phenotypes for the Working Dogs (특수 목적견으로서의 품성 및 능력 관련 유전자들에 관한 생물정보학적 분석)

  • Kwon, Yun-Jeong;Eo, Jungwoo;Choi, Bong-Hwan;Choi, Yuri;Gim, Jeong-An;Kim, Dahee;Kim, Tae-Hun;Seong, Hwan-Hoo;Kim, Heui-Soo
    • Journal of Life Science
    • /
    • v.23 no.11
    • /
    • pp.1325-1335
    • /
    • 2013
  • Working dogs, such as rescue dogs, military watch dogs, guide dogs, and search dogs, are selected by in-training examination of desired traits, including concentration, possessiveness, and boldness. In recent years, genetic information has been considered to be an important factor for the outstanding abilities of working dogs. To characterize the molecular features of the canine genes related to phenotypes for working dogs, we investigated the 24 previously reported genes (AR, BDNF, DAT, DBH, DGCR2, DRD4, MAOA, MAOB, SLC6A4, TH, TPH2, IFT88, KCNA3, TBR2, TRKB, ACE, GNB1, MSTN, PLCL1, SLC25A22, WFIKKN2, APOE, GRIN2B, and PIK3CG) that were categorized to personality, olfactory sense, and athletic/learning ability. We analyzed the chromosomal location, gene-gene interactions, Gene Ontology, and expression patterns of these genes using bioinformatic tools. In addition, variable numbers of tandem repeat (VNTR) or microsatellite (MS) polymorphism in the AR, MAOA, MAOB, TH, DAT, DBH, and DRD4 genes were reviewed. Taken together, we suggest that the genetic background of the canine genes associated with various working dog behaviors and skill performance attributes could be used for proper selection of superior working dogs.