• Title/Summary/Keyword: Gene Regulation

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STUDY CYTOCHROME P450IA1 GENE EXPRESSION BY RTPCR.

  • Lee, Soo-Young;Yhun Y. Sheen
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.128-128
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    • 1995
  • To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

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Legislation on Genetic Diagnosis: Comparison of South Korea and Germany - With Focus on the Application and Communication Structure -

  • Kim, Na-Kyoung
    • Development and Reproduction
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    • v.19 no.2
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    • pp.111-118
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    • 2015
  • This article explores the questions regarding PND and PID, especially the concrete legal conditions for the justification of PND and PID. As such, the German law stipulating PND and PID in a very concrete and detailed manner is introduced and explained in comparison with the corresponding South Korean law. The South Korean Bioethics and Biosafety Act (BBA) stipulates various types of gene testing and does not demonstrate a delicate sense of each type of gene testing. In contrast to the South Korean regulation, in Germany, there exist specific regulations for genetic counseling. Especially in the case of PND, GEKO stipulates the process of genetic counseling very concretely, based on GenDG. In the case of PND and PID, it is important that the people concerned understand the meaning of testing in various angles, and restructuralize it by combining it with their own values as the diagnosis is directly combined with pregnancy/abortion, which influences the whole life of a woman (and her partner). In this context, the South Korean BBA needs to be amended as soon as possible. The sections on informed consent also need to be amended to make them more concrete. Furthermore, guidelines for concretizing the regulation of BBA need to be continuously formulated and developed.

Effects of Mutations in the Regulatory Region on Transcriptional Regulation of glpD Gene

  • Jeong, Hee-Tae;Choi, Yong-Lark;Chung, Soo-Yeol;Chung, Chung-Han
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.245-249
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    • 1995
  • Expression of the adjacent but divergently transcribed glpD and glpE gene is positively regulated by cAMP-CRP. In this study, we constructed several mutants in which a CRP-binding site is placed at different distances upstream of the glpD promoter. The effect of the spacer length on transcription activation by cAMP-CRP was tested in vivo by $\beta$-galactosidase. The cAMP-CRP complex activated transcription from glpD when bound at a number of positions, all of which lay on the same face of the DNA helix, although the degree of activation varied with the length of the spacer. By contrast, the insertion of spacer length with non-integral turns of the DNA helix extremely inhibited the activation of transcription. The observed transcription activation by cAMP of the glpD promoter was influenced by the distance between the CRP binding site and the transcription start point.

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Effects of Five Fractions of Artemisia capillaris THUNB on $TGF{\beta}1-induced$ Apoptosis in HepG2 Cells (인진분획물이 인체간세포의 $TGF{\beta}1-induced$ Apoptosis에 미치는 영향)

  • 이지현;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.21 no.1
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    • pp.53-61
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    • 2000
  • Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

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Effect of Saliva miltiorrhiza Bunge on Antimicrobial Activity and Resistant Gene Regulation against Methicillin-Resistant Staphylococcus aureus (MRSA)

  • Lee, Ji-Won;Ji, Young-Ju;Lee, Syng-Ook;Lee, In-Seon
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.350-357
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    • 2007
  • This study was conducted in an effort to evaluate the antimicrobial activity and antibiotic-resistant gene regulation from Saliva miltiorrhiza Bunge on methicillin-resistant Staphylococcus aureus (MRSA). A variety of solvent fractions and methanol extracts of S. miltiorrhiza Bunge were tested in order to determine its antimicrobial activities against S. aureus and MRSA. As a result, the hexane fraction of S. miltiorrhiza Bunge evidenced the highest levels of antimicrobial activity against S. aureus and MRSA. The MICs of the hexane fraction against various MRSA specimens were $64. The hexane fraction evidenced inhibitory effects superior to those of the chloroform fraction. The results showed inhibition zones of hexane (16 mm) and chloroform (14 mm) fractions against MRSA KCCM 40511 at $1,000{\mu}g/disc$. The hexane and chloroform fractions inhibited the expression of the resistant genes, mecA, mecR1, and femA in mRNA. Moreover, the results of Western blotting assays indicated that the hexane and chloroform fractions inhibited the expression of the resistant protein, PBP2a. These results reveal that the hexane and chloroform fractions of S. miltiorrhiza Bunge may prove to be a valuable choice for studies targeted toward the development of new antimicrobial agents.

Cloning, DNA Sequence Determination, and Analysis of Growth-Associated Expression of the sodF Gene Coding for Fe- and Zn-Containing Superoxide Dismutase of Streptomyces griseus

  • Kim, Ju-Sim;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.700-706
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    • 2000
  • Iron- and zinc-containing superoxide dismutase (FeZnSOD) and nickel-containing superoxide dismutase (NiSOD) are cytoplamic enzymes in Streptomyces griseus. The sodF gene coding for FeZnSOD was cloned from genomic Southern hybridization analysis with a 0.5-kb DNA probe, which was PCR-amplified with facing primers corresponding to the N-terminal amino acid of the purified FeZnSOD of S. griseus and a C-terminal region which is conserved among bacterial FeSODs and MnSODs. The sodF open reading frame (ORF) was comprised of 213 amino acid (22,430 Da), and the deduced sequence of the protein was highly homologous (86% identity) to that of FeZnSOD of Streptomyces coelicolor. The FeZnSOD expression of exponentially growing S. griseus cell was approximately doubled as the cell growth reached the early stationary phase. The growth-associated expression of FeZnSOD was mainly controlled at the transcriptional level, and the regulation was exerted through the 110 bp regulatory DNA upstream from the ATG initiation codon of the sodF gene.

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DNA Double-Strand Breaks Serve as a Major Factor for the Expression of Arabidopsis Argonaute 2

  • Lee, Sungbeom;Chung, Moon-Soo;Lee, Gun Woong;Chung, Byung Yeoup
    • Journal of Radiation Industry
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    • v.10 no.4
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    • pp.243-248
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    • 2016
  • Argonaute 2 (AtAGO2) is a well characterized effector protein in Arabidopsis for its functionalities associated with DNA double-strand break (DSB)-induced small RNAs (diRNAs) and for its inducible expression upon ${\gamma}$-irradiation. However, its transcriptional regulation depending on the recovery time after the irradiation and on the specific response to DSBs has been poorly understood. We analyzed the 1,313 bp promoter sequence of the AtAGO2 gene ($1.3kb_{pro}$) to characterize the transcriptional regulation of AtAGO2 at various recovery times after ${\gamma}$-irradiation. A stable transformant harboring $1.3kb_{pro}$ fused with GUS gene showed that the AtAGO2 is highly expressed in response to ${\gamma}$-irradiation, after which the expression of the gene is gradually decreased until 5 days of DNA damage recovery. We also confirm that the AtAGO2 expression patterns are similar to that of ${\gamma}$-irradiation after the treatments of radiomimetic genotoxins (bleomycin and zeocin). However, methyl methanesulfonate and mitomycin C, which are associated with the inhibition of DNA replication, do not induce the expression of the AtAGO2, suggesting that the expression of the AtAGO2 is closely related with DNA DSBs rather than DNA replication.