• Title/Summary/Keyword: Inducible promoter

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MODULATION OF THE ACTIVITY OF PRO-INFLAMMATORY ENZYMES, COX-2 AND iNOS, BY CHRYSIN DERIVATIVES

  • Cho, Hee-Yeong;Yun, Cheol-Won;Kong, Jae-Yang;Kim, Kyoung-Soon;Park, You-Mie;Lee, Sang-Hyun;Kim, Bak-Kwang
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.286.1-286.1
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    • 2003
  • Chrysin, a natural flavone compound contained in plants. has anti-inflammatory activity. Its anti-inflammatory effect has been previously explained in part by the suppression of promoter activities of inducible pro-inflammatory enzymes (cyclooxygenase-2 (COX-2) and inducible nitrogen synthase (iNOS)). Nitrate production triggered by the activation of lipopolysaccharides (LPS) was most highly suppressed by the treatment of chrysin, follwed by 5-hydroxy-7-methoxyflavone (Ch-2), 5,7-diacetylflavone (Ch-4) in cultured Raw 264.7 cells. (omitted)

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Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.49-55
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    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.

Construction of a Reporter Strain Pseudomonas putida for the Detection of Oxidative Stress Caused by Environmental Pollutants

  • Lee Yun-Ho;Ahn Eun-Young;Park Sung-Su;Madsen Eugene L.;Jeon Che-Ok;Park Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.386-390
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    • 2006
  • A green fluorescent protein-based Pseudomonas putida reporter was successfully constructed and shown to be capable of detecting oxidative stress. In this whole-cell reporter, the promoter of the paraquat-inducible ferredoxin-$NADP^+$ reductase (fpr) was fused to a promoterless gfp gene on a broad-host-range promoter probe vector. Pseudomonas putida KT2440 harboring this reporter plasmid exhibited an increased level of gfp expression in the presence of redox-cycling agents (paraquat and menadione), hydrogen peroxide, and potential environmental pollutant chemicals such as toluene, paint thinner, gasoline, and diesel. Induction of fpr in the presence of these chemicals was confirmed using Northern blot analysis.

Auxin Effects on Symptom Development of Beet Curly Top Virus Infected Arabidopsis thaliana

  • Lee, Suk-Chan
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.249-256
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    • 1996
  • Beet curly top virus is the DNA virus that is providing useful for basic studies of the infection of Arabidopsis thaliana with viral host and provides a system for studying both resistance and the molecular basis of symptom development. An importnat aspect of symptom development observed in BCTV-infected A. thaliana (ecotype Sei-O) was the induction of cell division on phloem and surrounding cortex cells. Analysis of the expression of GUS reporter gene activity in transgenic plants containing constructs with promoter of the auxin-inducible saur gene showed that saur promoter activity was induced concomitantly in symptomatic tissues at the inflorescence shoot tips of the transgenic lines. The auxin sensitivity tests showed that hypersusceptible ecotype, Sei-O produced more amounts of callus than susceptible ecotype, Col-O. These studies indicated that changes in auxin concentration were involved in the induction of cell division in BCTV-infected plants and clearly demonstrated that there was a strong correlation between auxin-induced gene expression and the activation of cell division.

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Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

  • Chung, Injae
    • Toxicological Research
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    • v.16 no.2
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    • pp.95-100
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    • 2000
  • In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

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Regulation of the sufABCDSE Operon by Fur

  • Lee, Joon-Hee;Yeo, Won-Sik;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.109-114
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    • 2003
  • A promoter that is inducible by paraquat and menadione, the superoxide generators, independently of soxRS has been found in front of the sufABCDSE operon in Escherichia coli. Based on the observation that SufA is a holomog of IscA that functions in the assembly of iron sulfur cluster and the sufA promoter (sufAp) contains a putative Fur-binding consensus, we investigated whether this gene is regulated by Fur, a ferric uptake regulator, When examined in several sufAp-lacZ chromosomal fusion strains, sufAp was induced by EDTA, an iron chelator and a well-known Fur-inducer, The basal level of sufA expression increased dramatically in fur mutant, suggesting repression of sufAp by Fur. The derepression in fur mutant and EDTA-induction of sufA expression required nucleotides up to -61, where a putative Fur box is located. Purified Fur protein bound to the DNA fragment containing the putative Fur box between -35 and -10 promoter elements. The regulation by Fur and menadione induction of sufAp acted independently. The rpoS mutation increased sufA induction by menadione, suggesting that the stationary sigma factor RpoS acts negatively on sufA induction.

Construction of a Bile-responsive Expression System in Lactobacillus plantarum

  • Chae, Jong Pyo;Pajarillo, Edward Alain;Hwang, In-Chan;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.13-22
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    • 2019
  • This study aimed to develop a bile-responsive expression system for lactobacilli. The promoters of four genes, encoding phosphoenolpyruvate-dependent sugar phosphotransferase (mannose-specific), L-lactate dehydrogenase (LDH), HPr kinase, and D-alanine-D-alanine ligase, respectively, which were highly expressed by bile addition in Lactobacillus johnsonii PF01, were chosen. Each promoter was amplified by polymerase chain reaction and fused upstream of the ${\beta}$-glucuronidase gene as a reporter, respectively. Then, these constructs were cloned into E. coli-Lactobacillus shuttle vector pULP2, which was generated by the fusion of pUC19 with the L. plantarum plasmid pLP27. Finally, the constructed vectors were introduced into L. plantarum for a promoter activity assay. The LDH promoter showed the highest activity and its activity increased 1.8-fold by bile addition. The constructed vector maintained in L. plantarum until 80 generations without selection pressure. A bile-responsive expression vector, $pULP3-P_{LDH}$, for Lactobacillus spp. can be an effective tool for the bile-inducible expression of bioactive proteins in intestine after intake in the form of fermented dairy foods.

Cell growth and GFP expression in E. coli BL21 and W3110 under coexpression of Vitreoscilla hemogobin

  • Gang, Dong-Gyun;Kim, Yeon-Gyu;Cha, Hyeong-Jun
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.754-757
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    • 2001
  • Expression of the vhb gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been used to improve recombinant cell growth and enhance product formation under microaerobic conditions because of its ability to enhance oxygen use. We coexpressed GFP and VHb in Escherichia coli BL21 and W3110, and compared with GFP control which was not expressed VHb. We used nar oxygen-dependent inducible promoter for VHb expression. The GFP amounts in E. coli expressed VHb was about five fold higher than in the control Fluorescence intensity was increased about two fold.

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Allantoin 분해 유전자들의 발현 유도에 관여하는 세가지 요소 (UAS, URS, UIS)

  • 유향숙
    • The Microorganisms and Industry
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    • v.14 no.1
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    • pp.12-16
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    • 1988
  • Allantoin 분해 유전자들중 highly inducible 한 DAL7, DUR1,2및 constitutive한 DAL5 gene의 promoter를 deletion 방법에 의해 발현에 필요한 최소 DNA seqyence 부위를 정한후 이 DNA seqyence를 다시 oligonucleotide 합성방법에 의해 합성하여 Cyc 1-LacZ expression vector에 삽입하여 효모내에서 LacZ의 발현이 삽입한 DNA sequence에 의해 영향을 받는 정도를 측정하여 (.betha.-galactosidase activity) deletion 방법에 의해 결정한 이 DNA dequence들이 직접 발현유도에 관여하는가를 조사하였다.

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