• Title/Summary/Keyword: IPTG

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A VoIP Traffic Generator for Simulating Call Processing in IP Contact Center Systems (IP 컨택센터에서 통화 처리 모의 실험을 위한 VoIP 트래픽 생성기)

  • Kim Soo-Hee;Jung In-Hwan
    • Proceedings of the Korea Information Processing Society Conference
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    • 2006.05a
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    • pp.1239-1242
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    • 2006
  • 본 논문에서는 IP 컨택센터에서 통화 처리 모의실험을 위한 VoIP 트래픽 발생기를 설계하고 구현한다. 구현된 트래픽 발생기(VoIPTG)는 H.323과 SIP 기반의 VoIP 콜 생성과 음성코덱(G.711, G.723.1등)을 사용하는 RTP 트래픽을 발생시킴으로써 다수의 대화자들이 음성 통화하는 상황을 모의실험을 할 수 있도록 해준다. VoIPTG를 이용하면 H.323 또는 SIP 세션 제어 프로토콜 선택, 사용자(call)수 변화, 시간 변화, 음성코덱의 선택 등 여러 가지 조합을 통해 다양한 모의실험 환경을 연출 할 수 있다. 이러한 트래픽 발생기는 IP 기반 컨택센터의 전반적인 기능 검사 및 성능평가를 위해 유용하게 사용될 수 있으며, 특히 IP 기반 녹취 시스템의 성능 평가를 위해서 필수적이다.

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Pseudoalteromonas carrageenovora 유래 Arylsulfatase의 cloning과 재조합 E. coli에서 과발현

  • Im, Jae-Myeong;Kim, Hyeong-Rak;Kim, Seong-Gu;Nam, Su-Wan
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.571-575
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    • 2003
  • A marine aerobic Gram-negative bacterium, Pseudoalteromonas carrageenovora, has been blown to hydrolyze carrageenans, the sulfated galactans of red algae, and to desulfate oligo kappa-carrageenans. Recently, the gene encoding arylsulfatase (aryl-sulfate sulfohydrolase, E.C.3.1.6.1) of A. carrageenovora was cloned and the nucleotide sequence was reported. Enzymatic hydrolysis of sulfate groups in agaropectin simplifies the process of agarose preparation. In order to overproduce the enzyme, the arylsulfatase gene (astA, 984 bp ORF) from P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 was introduced into E, coli BL21(DE3), the transformant on LB plate containing IPTG showed the hydrolyzing activity for p-nitrophenyl sulfate. Most of arylsulfatase activity was found in the cell lysate, but at $50\;{\sim}\;5000\;{\mu}M$ IPTG concentration the activity was found both in the culture supernatant and the cell lysate. The molecular weight of the recombinant enzyme was estimated to be 34 kDa by SDS-PAGE.

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Overproduction of Bacillus macerans Cyclodextrin Glucanotransferase in E. coli by Coexpression of GroEL/ES Chaperone

  • Kwon, Mi-Jung;So-Lim Park;Sung-Koo Kim;Soo-Wan Nam
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.1002-1005
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    • 2002
  • The effects of GroEL/ES chaperone on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) in recombinant E. coli were investigated. The cgt gene and groEL/ES genes are under the control of T7 promoter and Pzt-1 promoter, respectively. The optimal concentrations of inducers, IPTG and tetracycline, were found to be 1.0 mM and 10 ng/ml, respectively. When tetracycline and IPTG were added at the early exponential phase (2h) and exponential phase (3h) of growth, respectively, about 1.5-fold increase of soluble CGTase activity and 1.6-fold increase of soluble CGTase protein were obtained. An SDS-PAGE analysis revealed that about $37.2\%$ of total CGTase protein was in the soluble fraction when GroEL/ES chaperone was overexpressed.

Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli

  • Park, Eun-Hee;Lee, Won-Heong;Jang, Mi-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1709-1713
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    • 2015
  • Sepiapterin is a precursor for the synthesis of tetrahydrobiopterin (BH4), which is a wellknown cofactor for aromatic amino acid hydroxylation and nitric oxide synthesis in higher mammals. In this study, a recombinant Escherichia coli BL21(DE3) strain harboring cyanobacterial guanosine 5’-triphosphate cyclohydrolase 1 (GCH1) and human 6-pyruvoyltetrahydropterin synthase (PTPS) genes was constructed to produce sepiapterin. The optimum conditions for T7 promoter–driven expression of GCH1 and PTPS were 30℃ and 0.1 mM isopropyl-β-D-thioglucopyranoside (IPTG). The maximum sepiapterin concentration of 88.1 ± 2.4 mg/l was obtained in a batch cultivation of the recombinant E. coli, corresponding to an 18-fold increase in sepiapterin production compared with the control condition (37℃ and 1 mM IPTG).

재조합 Escherichia coli 시스템을 이용한 폐흡충 cystein proteinase의 생산 연구

  • Hong, Seong-Hui;Lee, Gil-Hwan;Jeon, Hui-Jin;Hwang, Hyeon-A;Park, Seong-Ryeol;Hwang, Yeong-Bo;Park, Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.651-654
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    • 2000
  • A fermentation study of the recombinant Escherichia coli system has been tired to overproduce a recombinant Paragonimus westermani cysteine proteinase, rPwCP1. Using the modified LB media, main cultures and chemical induction with IPTG were carried out to examine the possibility of secretory protein overproduction at high cell density culture. As a result, the target protein of rPwCP1, purified by metal affinity chromatography and gel filtration, has been shown at 50.8 kDa on SDS-PAGE, and its final concentration turns out to be 350mg/L.

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Optimization of gamma-Aminobutyric Acid Bioconversion by Recombinant Escherichia coli (재조합 Escherichia coli를 이용한 gamma-Aminobutyric Acid 전환 반응 최적화)

  • Le Vo, Tam Dinh;Hong, Soon-Ho
    • KSBB Journal
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    • v.27 no.2
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    • pp.127-130
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    • 2012
  • In this study, the effects of pH, temperature, IPTG concentration and substrate (MSG) concentration on gamma-aminobutyric acid (GABA) production in engineered Escherichia coli were investigated. Glutamate decarboxylase and glutamate/GABA antiporter were overexpressed in GABA aminotransferase knock-out strain for GABA production. The result of optimization study showed the GABA bioconversion was optimized at pH 3.5, $30^{\circ}C$, 0.5 mM IPTG, 10 g/L MSG. At this condition, 5.23 g/L of final GABA concentration of was achieved from 10 g/L of MSG, which corresponded to a GABA yield of 85.77%.

Optimization of Culture Conditions for Production of Helicobacter pylori Adhesin Protein Genetically Linked to Cholera Toxin A2B in Escherichia coli JM101

  • Kim, Byung-Oh;Pyo, Suh-Kneung
    • Biomolecules & Therapeutics
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    • v.9 no.3
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    • pp.162-166
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    • 2001
  • Helicobacter pylori is a major cause of gastric-associated diseases. In our previous study, the Adhesin/CTXA2B was expressed as insoluble recombinant chimeric protein derived from the H. pylori adhesin genetically coupled to CTXA2B subunit in Escherichia coli. Since it is very important to optimize IPTG concentration, culture temperature and composition of medium to maximize cell growth and productivity, these conditional growth factors were determined for increasing the productivity of the expressed Adhesin/CTXA2B chimeric protein in Escherichia coli JM101 carrying pTEDhpa/ctxa2b. Our data demonstrate that optimal medium for increased production of chimeric protein was a YCP/Glu medium composed of 2% yeast extract, 1% casamino acid, phosphate solution [0.3% $KH_2P0_4$, 0.4% $Na_2HP0_4$, 0.25% ($NH_4)_2HPO_4$], and 0.5% glucose. In addition, optimal concentration of IPTG was 1 mM and culture temperature, $37^{\circ}C$.

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Extracellular Production of Alpha-Interferon by Recombinant Escherichia coli: PartIV. Effects of Ampicillin and an Inducer on the Production of Alpha-Interferon and Plasmid Stability (유전자 재조합 대장균을 사용한 Alpha-interferon의 생산과 분비;제4부. Ampicillin 및 Inducer의 Alpha-interferon의 생산과 Plasmid 안정성에 미치는 영향)

  • 노갑수;최차용
    • KSBB Journal
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    • v.6 no.1
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    • pp.9-14
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    • 1991
  • We studied the production and excretion of alpha-interferon in recombinant Escherichia coli harboring plasmid pIF-III-B, which carries alpha-interferon gene under the control of lipoprotein and lacUV5 promoter, and lac operator. Basically, the effects of concentrations of ampicillin and an inducer, IPTG, for the expression of the cloned gene, on the productions of alpha-interferon and plasmid stability were studied. The highest production of alpha-interferon was observed at 50 mg/1 of ampicillin concentration and 0.5 mM of IPTG. The plasmid pIF-III-B was maintained very stably in medium with ampicillin but segregated rapidly in medium without ampicillin. Also, the plasmid was segregated more rapidly in medium with an inducer higher than 0.5 mM.

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Effects of a Processing Inhibitor on the Overproduction of Plasmid Encoded ${\beta}$-lactamase in E. coli (대장균 ${\beta}$-lactamase의 대량생산시 Processing Inhibitor의 영향)

  • 홍원경;김은기
    • KSBB Journal
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    • v.6 no.1
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    • pp.111-114
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    • 1991
  • The effects of the precursor processing inhibitor, carbonylcyanide-chlorophenyl hydrazone(CCCP), are investigated on the production of soluble ${\beta}-lactamase$ the formation of the inclusion body when ${\beta}-lactamase$ is overproduced by induction with isopropyl thiogalactoside(IPTG). When cells are treated by CCCP, more soluble ${\beta}-lactamase$ is produced. In this case, no difference in the amount of inclusion body is observed.

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