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Molecular Cloning of Novel Genes Specifically Expressed in Snailfish, Liparis tanakae (꼼치, Liparis tanakae에서 특이하게 발현되는 새로운 유전인자의 검색)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.67-77
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    • 2000
  • Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

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Effects Unripe and Ripe Rubus coreanus Miquel on Peritoneal Macrophage Gene Expression Using cDNA Microarray Analysis (미숙과와 성숙과 복분자의 섭취가 복강 Macrophages의 유전자 발현에 미치는 영향)

  • Lee, Jung Eun;Cho, Soo-Muk;Kim, Jin;Kim, Jung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1552-1559
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    • 2013
  • Rubus coreanus Miquel (RCM) has been used as one of the Korean traditional medicines for prostate health. In addition, recent studies have reported that RCM reduced chronic inflammatory diseases such as cancer, and rheumatoid arthritis. Therefore, in this study, we investigated the effects of unripe and ripe RCM on inflammationrelated gene expressions in LPS-stimulated mouse peritoneal macrophages. Mice were fed with 2% unripe RCM (U2), 10% unripe RCM (U10), 2% ripe RCM (R2), and 10% ripe RCM (R10) for 8 weeks. Peritoneal macrophages were isolated and stimulated with LPS then proinflammatory mediators (TNF-${\alpha}$, IL-$1{\beta}$, and IL-6), and prostaglandin E2 ($PGE_2$) productions were assessed. Moreover, gene expression profiles were analyzed by cDNA microarray method. Unripe and ripe RCM significantly reduced TNF-${\alpha}$ production but only unripe RCM decreased IL-$1{\beta}$ and IL-6 production. RCM intake significantly reduced inflammatory-related gene expressions such as arachidonate 5-lipoxygenase, interleukin 11, and nitric oxide synthase 2. Furthermore, unripe and ripe RCM significantly decreased ceruloplasmin, tissue plasminogen activator, thrombospondin 1, and vascular endothelial growth factor A expression which modulates symptoms of chronic inflammatory diseases. RCM intake also significantly increased hypoxia inducible factor 3, alpha which is the negative regulators of hypoxia-inducible gene expression. Furthermore, only unripe RCM reduced chemokine (C-C motif) ligand 8, chemokine (C-X-C motif) ligand 14, and phospholipase A2 expression. In this study, we showed that RCM had anti-inflammatory effects by suppression of pro-inflammatory mediator expressions and may reduce chronic inflammatory disease progress through regulation of gene expressions. These findings suggest that RCM might be used as a potential functional material to reduce chronic inflammatory responses.

A Study on History and Archetype Technology of Goli-su in Korea (한국 고리수의 역사와 원형기술의 복원 연구)

  • Kim, Young-ran
    • Korean Journal of Heritage: History & Science
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    • v.46 no.2
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    • pp.4-25
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    • 2013
  • Goli-su is the innovative special kind of the embroidery technique, which combines twining and interlacing skill with metal technology and makes the loops woven to each other with a strand. The loops floating on the space of the ground look like floating veins of sculpture and give people the feeling of the openwork. This kind of characteristic has some similarities with the lacework craft of Western Europe in texture and technique style, but it has its own features different from that of Western Europe. It mainly represents the splendid gloss with metallic materials in the Embroidered cloth, such as gold foil or wire. In the 10th century, early days of Goryo, we can see the basic Goli-su structure form of its initial period in the boy motif embroidery purse unearthed from the first level of Octagonal Nine-storied Pagoda of Woljeong-sa. In the Middle period of Joseon, there are several pieces of Goli-su embroidered relic called "Battle Flag of Goryo", which was taken by the Japanese in 1592 and is now in the Japanese temple. This piece is now converted into altar-table covers. In 18~19th century, two pairs of embroidered pillows in Joseon palace were kept intact, whose time and source are very accurate. The frame of the pillows was embroidered with Goli-su veins, and some gold foil papers were inserted into the inside. The triangle motif with silk was embroidered on the pillow. The stitch in the Needle-Looped embroidery is divided into three kinds according to comprehensive classification: 1. Goli-su ; 2. Goli-Kamgi-su ; 3. Goli-Saegim-su. From the 10th century newly establishing stage to the 13th century, Goli-su has appeared variational stitches and employed 2~3 dimensional color schemes gradually. According to the research of this thesis, we can still see this stitch in the embroidery pillow, which proves that Goli-suwas still kept in Korea in the 19th century. And in terms of the research achievement of this thesis, Archetype technology of Goli-su was restored. Han Sang-soo, Important Intangible Cultural Heritage No. 80 and Master of Embroidery already recreated the Korean relics of Goli-su in Joseon Dynasty. The Needle-Looped embriodery is the overall technological result of ancestral outstanding Metal craft, Twining and Interlacing craft, and Embroidery art. We should inherit, create, and seek the new direction in modern multi-dimensional and international industry societyon the basis of these research results. We can inherit the long history of embroidering, weaving, fiber processing, and expand the applications of other craft industries, and develop new advanced additional values of new dress material, fashion technology, ornament craft and artistic design. Thus, other crafts assist each other and broaden the expressive field to pursue more diversified formative beauty and beautify our life abundantly together.

Molecular Cloning, Characterization, and Expression Analysis of Chicken Δ-6 Desaturase

  • Kang, Xiangtao;Bai, Yichun;Sun, Guirong;Huang, Yanqun;Chen, Qixin;Han, Ruili;Li, Guoxi;Li, Fadi
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.1
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    • pp.116-121
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    • 2010
  • Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.

Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

  • Choi, Jae-Woong;Kim, Jong-Wook;Nguyen, Lap P.;Nguyen, Huu C.;Park, Eun-Mee;Choi, Dong Hwa;Han, Kang Min;Kang, Sang Min;Tark, Dongseob;Lim, Yun-Sook;Hwang, Soon B.
    • Molecules and Cells
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    • v.43 no.5
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    • pp.469-478
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    • 2020
  • Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP-1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.

In silico Design of Discontinuous Peptides Representative of B and T-cell Epitopes from HER2-ECD as Potential Novel Cancer Peptide Vaccines

  • Manijeh, Mahdavi;Mehrnaz, Keyhanfar;Violaine, Moreau;Hassan, Mohabatkar;Abbas, Jafarian;Mohammad, Rabbani
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5973-5981
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    • 2013
  • At present, the most common cause of cancer-related death in women is breast cancer. In a large proportion of breast cancers, there is the overexpression of human epidermal growth factor receptor 2 (HER2). This receptor is a 185 KDa growth factor glycoprotein, also known as the first tumor-associated antigen for different types of breast cancers. Moreover, HER2 is an appropriate cell-surface specific antigen for passive immunotherapy, which relies on the repeated application of monoclonal antibodies that are transferred to the patient. However, vaccination is preferable because it would stimulate a patient's own immune system to actively respond to a disease. In the current study, several bioinformatics tools were used for designing synthetic peptide vaccines. PEPOP was used to predict peptides from HER2 ECD subdomain III in the form of discontinuous-continuous B-cell epitopes. Then, T-cell epitope prediction web servers MHCPred, SYFPEITHI, HLA peptide motif search, Propred, and SVMHC were used to identify class-I and II MHC peptides. In this way, PEPOP selected 12 discontinuous peptides from the 3D structure of the HER2 ECD subdomain III. Furthermore, T-cell epitope prediction analyses identified four peptides containing the segments 77 (384-391) and 99 (495-503) for both B and T-cell epitopes. This work is the only study to our knowledge focusing on design of in silico potential novel cancer peptide vaccines of the HER2 ECD subdomain III that contain epitopes for both B and T-cells. These findings based on bioinformatics analyses may be used in vaccine design and cancer therapy; saving time and minimizing the number of tests needed to select the best possible epitopes.

Deoxynivalenol- and zearalenone-contaminated feeds alter gene expression profiles in the livers of piglets

  • Reddy, Kondreddy Eswar;Jeong, Jin young;Lee, Yookyung;Lee, Hyun-Jeong;Kim, Min Seok;Kim, Dong-Wook;Jung, Hyun Jung;Choe, Changyong;Oh, Young Kyoon;Lee, Sung Dae
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.595-606
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    • 2018
  • Objective: The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods: In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results: Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone $H_2B$ type 1, and serum amyloid A). Conclusion: In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.

Extraction and Indexing Representative Melodies Considering Musical Composition Forms for Content-based Music Information Retrievals (내용 기반 음악 정보 검색을 위한 음악 구성 형식을 고려한 대표 선율의 추출 및 색인)

  • Ku, Kyong-I;Lim, Sang-Hyuk;Lee, Jae-Heon;Kim, Yoo-Sung
    • The KIPS Transactions:PartD
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    • v.11D no.3
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    • pp.495-508
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    • 2004
  • Recently, in content-based music information retrieval systems, to enhance the response time of retrieving music data from large music database, some researches have adopted the indexing mechanism that extracts and indexes the representative melodies. The representative melody of music data must stand for the music itself and have strong possibility to use as users' input queries. However, since the previous researches have not considered the musical composition forms, they are not able to correctly catch the contrast, repetition and variation of motif in musical forms. In this paper, we use an index automatically constructed from representative melodies such like first melody, climax melodies and similarly repeated theme melodies. At first, we expand the clustering algorithm in order to extract similarly repeated theme melodies based on the musical composition forms. If the first melody and climax melodies are not included into the representative melodies of music by the clustering algorithm, we add them into representative melodies. We implemented a prototype system and did experiments on comparison the representative melody index with other melody indexes. Since, we are able to construct the representative melody index with the lower storage by 34% than whole melody index, the response time can be decreased. Also, since we include first melody and climax melody which have the strong possibility to use as users' input query into representative melodies, we are able to get the more correct results against the various users' input queries than theme melody index with the cost of storage overhead of 20%.

Creative Curiosity: Study of Alice Character in Lewis Caroll's Adventures of Alice in Wonderland (창조적 호기심 루이스 캐럴의 『이상한 나라의 앨리스의 모험』 연구)

  • Cho, Sungran
    • Cross-Cultural Studies
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    • v.41
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    • pp.299-320
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    • 2015
  • Lewis Carroll's Alice's Adventures in Wonderland expands scope of Children's Literature genre by introducing the discourse of pleasure as opposed to that of didactic discipline. Carroll's narrative is important, not only for children's literature, but also as a forerunner of post/modernism of James Joyce with its language play and linguistic invention. Its treatment of Alice's body change follows the motif of body transformation in myth and literature. Comparing "stasis" of Susan Sontag's character Alice (James) in her play Alice in Bed and "movement" of Carroll's Alice, this study explores the issues of woman's alienation and the dichotomy of mobility/immobility in reality and in their literary representations. Focusing on a female child's double alienation as woman and child, I argue Alice's Adventures in Wonderland is a counter-narrative of alternative bildungsroman. Alice gains her subjectivity through her adventure by power of language and story-telling. Through representation of the dream/adventure of two desiring sisters, Carroll's narrative exhibits subversion of social order and emergence of new order of "chaosmos" out of chaos. As a method of study, this study traces genealogy of "curiosity" in myth and literature as a motivating force that triggers adventure and argues "creative curiosity" is a dynamic energy propelling Alice's adventure.

Site-specific Dye-labeling of the Bacterial Cell Surface by Bioconjugation and Self-assembly (바이오접합과 자가결합을 이용한 박테리아 세포막의 위치 특이적 형광 표지)

  • Yang, I Ji;Lim, Sung In
    • Korean Chemical Engineering Research
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    • v.60 no.3
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    • pp.398-406
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    • 2022
  • The outer membrane of Gram-negative bacteria is the outermost layer of cellular environment in which numerous biophysical and biochemical processes are in action sustaining viability. Advances in cell engineering enable modification of bacterial genetic information that subsequently alters membrane physiology to adapt bacteria to specific purposes. Surface display of a functional molecule on the outer membranes is one of strategies that directs host cells to respond to a specific extracellular matter or stimulus. While intracellular expression of a functional peptide or protein fused to a membrane-anchoring motif is commonly practiced for surface display, the method is not readily applicable to exogenous or large proteins inexpressible in bacteria. Chemical conjugation at reactive groups naturally occurring on the membrane might be an alternative, but often compromises fitness due to non-specific modification of essential components. Herein, we demonstrated two distinct approaches that enable site-specific decoration of the outer membrane with a fluorescent agent in Escherichia coli. An unnatural amino acid genetically incorporated in a surface-exposed peptide could act as a chemoselective handle for bioorthogonal dye labeling. A surface-displayed α-helical domain originating from a part of a selected heterodimeric coiled-coil complex could recruit and anchor a green fluorescent protein tagged with a complementary α-helical domain to the membrane surface in a site- and hetero-specific manner. These methods hold a promise as on-demand tools to confer new functionalities on the bacterial membranes.