• 제목/요약/키워드: Systems Biotechnology

검색결과 1,501건 처리시간 0.023초

Optimized Expression, Purification, and Rapid Detection of Recombinant Influenza Nucleoproteins Expressed in Sf9 Insect Cells

  • Yoon, Sung-Jin;Park, Young-Jun;Kim, Hyun Ju;Jang, Jinwoo;Lee, Sang Jun;Koo, Sunwoo;Lee, Moo-Seung
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1683-1690
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    • 2018
  • Accurate and rapid diagnosis of influenza infection is essential to enable early antiviral treatment and reduce the mortality associated with seasonal and epidemic infections. Immunochromatography is one of the most common methods used for the diagnosis of seasonal human influenza; however, it is less effective in diagnosing pandemic influenza virus. Currently, rapid diagnostic kits for pandemic influenza virus rely on the detection of nucleoprotein (NP) or hemagglutinin (HA). NP detection shows higher specificity and is more sensitive than HA detection. In this study, we time-dependently screened expression conditions, and herein report optimal conditions for the expression of recombinant nucleoprotein (rNP), which was 48 h after infection. In addition, we report the use of the expressed rNP in a rapid influenza diagnostic test (SGT i-flex Influenza A&B Test). We constructed expression vectors that synthesized rNP (antigen) of influenza A and B in insect cells (Sf9 cells), employed the purified rNP to the immunoassay test kit, and clearly distinguished NPs of influenza A and influenza B using this rapid influenza diagnostic kit. This approach may improve the development of rapid test kits for influenza using NP.

EST Knowledge Integrated Systems (EKIS): An Integrated Database of EST Information for Research Application

  • Kim, Dae-Won;Jung, Tae-Sung;Choi, Young-Sang;Nam, Seong-Hyeuk;Kwon, Hyuk-Ryul;Kim, Dong-Wook;Choi, Han-Suk;Choi, Sang-Heang;Park, Hong-Seog
    • Genomics & Informatics
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    • 제7권1호
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    • pp.38-40
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    • 2009
  • The EST Knowledge Integrated System, EKIS (http://ekis.kribb.re.kr), was established as a part of Korea's Ministry of Education, Science and Technology initiative for genome sequencing and application research of the biological model organisms (GEAR) project. The goals of the EKIS are to collect EST information from GEAR projects and make an integrated database to provide transcriptomic and metabolomic information for biological scientists. The EKIS constitutes five independent categories and several retrieval systems in each category for incorporating massive EST data from high-throughput sequencing of 65 different species. Through the EKIS database, scientists can freely access information including BLAST functional annotation as well as Genechip and pathway information for KEGG. By integrating complex data into a framework of existing EST knowledge information, the EKIS provides new insights into specialized metabolic pathway information for an applied industrial material.

Insights into Enzyme Reactions with Redox Cofactors in Biological Conversion of CO2

  • Du-Kyeong Kang;Seung-Hwa Kim;Jung-Hoon Sohn;Bong Hyun Sung
    • Journal of Microbiology and Biotechnology
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    • 제33권11호
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    • pp.1403-1411
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    • 2023
  • Carbon dioxide (CO2) is the most abundant component of greenhouse gases (GHGs) and directly creates environmental issues such as global warming and climate change. Carbon capture and storage have been proposed mainly to solve the problem of increasing CO2 concentration in the atmosphere; however, more emphasis has recently been placed on its use. Among the many methods of using CO2, one of the key environmentally friendly technologies involves biologically converting CO2 into other organic substances such as biofuels, chemicals, and biomass via various metabolic pathways. Although an efficient biocatalyst for industrial applications has not yet been developed, biological CO2 conversion is the needed direction. To this end, this review briefly summarizes seven known natural CO2 fixation pathways according to carbon number and describes recent studies in which natural CO2 assimilation systems have been applied to heterogeneous in vivo and in vitro systems. In addition, studies on the production of methanol through the reduction of CO2 are introduced. The importance of redox cofactors, which are often overlooked in the CO2 assimilation reaction by enzymes, is presented; methods for their recycling are proposed. Although more research is needed, biological CO2 conversion will play an important role in reducing GHG emissions and producing useful substances in terms of resource cycling.

Anammox Bacteria Enrichment in Upflow Anaerobic Sludge Blanket (UASB) Reactor

  • Thuan Tran-Hung;Jahng Deok-Jin;Jung Jin-Young;Kim Dong-Jin;Kim Won-Kyoung;Park Young-Joo;Kim Ji-Eun;Ahn Dae-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권5호
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    • pp.345-351
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    • 2004
  • We investigated the anaerobic ammonium oxidation (anammox) reaction in a lab-stale upflow anaerobic sludge blanket (UASB) reactor. Our aim was to detect and enrich the organisms responsible for the anammox reaction using a synthetic medium that contained low concentrations of substrates (ammonium and nitrite). The reactor was inoculated with granular sludge collected from a full-scale anaerobic digestor used for treating brewery wastewater The experiment was performed during 260 days under conditions of constant ammonium concentration ($50\;mg\;NH_4^+-N/L$) and different nitrite concentrations ($50{\~}150\;mg\;NO_2-N/L$). After 200 days, anammox activity was observed in the system. The microorganisms involved in this anammox reaction were identified as Candidatus B. Anammoxidans and K. Stuttgartiensis using fluorescence in situ hybridization (FISH ) method.

SETDB1 regulates SMAD7 expression for breast cancer metastasis

  • Ryu, Tae Young;Kim, Kwangho;Kim, Seon-Kyu;Oh, Jung-Hwa;Min, Jeong-Ki;Jung, Cho-Rok;Son, Mi-Young;Kim, Dae-Soo;Cho, Hyun-Soo
    • BMB Reports
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    • 제52권2호
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    • pp.139-144
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    • 2019
  • Breast cancer (BRC) is the most invasive cancer in women. Although the survival rate of BRC is gradually increasing due to improved screening systems, development of novel therapeutic targets for inhibition of BRC proliferation, metastasis and recurrence have been constantly needed. Thus, in this study, we identified overexpression of SETDB1 (SET Domain Bifurcated 1), a histone methyltransferase, in RNA-seq data of BRC derived from TCGA portal. In Gene Ontology (GO) analysis, cell migration-related GO terms were enriched, and we confirmed down-regulation of cell migration/invasion and alteration of EMT /MET markers after knockdown of SETDB1. Moreover, gene network analysis showed that SMAD7 expression is regulated by SETDB1 levels, indicating that up-regulation of SMAD7 by SETDB1 knockdown inhibited BRC metastasis. Therefore, development of SETDB1 inhibitors and functional studies may help develop more effective clinical guidelines for BRC treatment.

The Production of Transgenic Livestock and Its Applications

  • Han, Y. M;Lee, K. K.
    • 한국가축번식학회지
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    • 제23권4호
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    • pp.381-391
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    • 1999
  • During the last 20 years, transgenic animal technology has provided revolutionary new opportunities in many aspects of agriculture and biotechnology. Several gene delivery systems including pronuclear injection, retroviral vectors, sperm vectors, and somatic cell cloning have developed for making transgenic animals. In the future major improvements in transgenic animal generation will be mainly covered by somatic cell cloning technology. Many factors affecting integration frequency and expression of the transgenes should be overcome to facilitate the industrial applications of transgenic technology. Transgenic animal technology has settled down in some areas of the biotechnology, especially the mass production of valuable human proteins and xenotransplantation. In the 21st century animal biotechnology will further contribute to welfare of human being.

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Biosynthesis of Two Flavones, Apigenin and Genkwanin, in Escherichia coli

  • Lee, Hyejin;Kim, Bong Gyu;Kim, Mihyang;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1442-1448
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    • 2015
  • The flavonoid apigenin and its O-methyl derivative, genkwanin, have various biological activities and can be sourced from some vegetables and fruits. Microorganisms are an alternative for the synthesis of flavonoids. Here, to synthesize genkwanin from tyrosine, we first synthesized apigenin from p-coumaric acid using four genes (4CL, CHS, CHI, and FNS) in Escherichia coli. After optimization of different combinations of constructs, the yield of apigenin was increased from 13 mg/l to 30 mg/l. By introducing two additional genes (TAL and POMT7) into an apigenin-producing E. coli strain, we were able to synthesize 7-O-methyl apigenin (genkwanin) from tyrosine. In addition, the tyrosine content in E. coli was modulated by overexpressing aroG and tyrA. The engineered E. coli strain synthesized approximately 41 mg/l genkwanin.

Phenylalanyl-2-Sulfanilylglycine as Substrate for Leucine Aminopeptidase Assay

  • Hwang, Se-Young;Cho, Suk-Young;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • 제5권6호
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    • pp.319-323
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    • 1995
  • A chromogenic mimic of phenlyalanyl-dipeptide, L-phenylalanyl-L-2-sulfanilylglycine (PSG), was synthesized and examined for its usability in leucine aminopeptidase (LAP) assay. The enzyme activity was easily determined by measuring the amount of diazotized adduct of sulfanilic acid released upon hydrolysis of PSG ($\varepsilon^{420}$=18,000/M/cm). Under the experimental conditions employed, PSG showed a Km of 0.063 mM and a Kcat of 1683/min, assessable less than 0.1 $\mu$ g of LAP per milliliter. And the presence of aminopeptidase M (APM) was suggested to be negligible in LAP assay. This novel assay can circumvent the occasional yellow background in biological systems, i.e., serums, etc..

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Anti-angiogenic activity of wilfoside glycosides isolated from Cynanchum wilfordii

  • Kim, Myoung-Sook;Baek, Jin-Hyen;Lee, Hyo-Jong;Hwang, Bang-Yeon;Kim, Se-Eun;Lee, Jung-Joon;Kim, Kyu-Won
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.164.1-164.1
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    • 2003
  • Wm was isolated from Cynanchum wilfordii (Asclepiadaceae) as a mixture form of polypregnane glycosides that included wilfoside K1N and wilfoside C1N. In the present study, we investigated the anti-angiogenic effect of wilfoside glycosides using in vivo and in vitro assay systems. We first demonstrated that concentrated conditioned media obtained from Wm-treated HepG2 human hepatoblastoma cells blocked the angiogenic activation of Wm-untreated concentrated conditioned media, suggesting that Wm may have an inhibitory effect on tumor-induced angiogenesis. (omitted)

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Identification of matrix metalloproteinases secreted by human hepatocarcinoma HepG2 cells

  • Lee, Young Jae;Kim, Keun Cheon;Lim, Jeong Mook;Lee, Seung Tae
    • 한국동물생명공학회지
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    • 제37권1호
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    • pp.62-66
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    • 2022
  • To date, the development of anticancer drugs has been conducted using two-dimensional (2D) cell culture systems. However, since cancer cells in the body are generated and developed in three-dimensional (3D) microenvironments, the use of 2D anticancer drug screening can make it difficult to accurately evaluate the anticancer effects of drug candidates. Therefore, as a step towards developing a cancer cell-friendly 3D microenvironment based on a combination of vinylsulfone-functionalized polyethylene glycol (PEG-VS) with dicysteine-containing crosslinker peptides with an intervening matrix metalloproteinase (MMP)-specific cleavage site, the types of MMPs secreted from human hepatocarcinoma HepG2 cells, a representative cancer cell, were analyzed transcriptionally and translationally. MMP3 was confirmed to be the most highly expressed protease secreted by HepG2 cells. This knowledge will be important in the design of a crosslinker necessary for the construction of PEG-based hydrogels customized for the 3D culture of HepG2 cells.