• Title/Summary/Keyword: synthetic gene

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Effect of an Anabolic Steroid, Nandrolone Decanoate, on Aquaporin 1 and 9 Gene Expression in the Rat Epididymis

  • Seo, Hee-Jung;Kang, Hyo-Jin;Choi, In-Ho;Cheon, Yong-Pil;Lee, Ki-Ho
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.55-61
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    • 2009
  • The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

Induction of Erythromycin by Virginiamycin Inducing Factor, Virginiae Butanolide C (Virginiamycin 생합성 유도인자 Virginiae Butanolide C에 이한 Erythromycin 생산 유도)

  • Kim, Hyun-Soo;Seong, Lim-Shik
    • KSBB Journal
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    • v.14 no.6
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    • pp.682-687
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    • 1999
  • Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the production of virginiamycin in Streptomyces virginiae. In order to investigate the function of VB-C as inducer in other strains, Streptomyces erythraeus was used as a test strain(parent). VB-C binding receptor gene was introduced into S. erythraeus(transformant) and the production of VBs and specific VB-C binding protein were analysed in parent and transformant. When 300ng/ml of the synthetic VB-C was added at 0, 20, 44 h cultivation of the parent and at 44 h cultivation of the transformant, the initial production times a antibiotics were shortened by more than 8 and 6 h, respectively. The transformant showed strong antibiotic activity against B. subtilis. These results suggest that the VB-C might have an ability to induce the production of secondary metabolites in S. erythraeus.

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Transcriptional Regulation of the AP-1 and Nrf2 Target Gene Sulfiredoxin

  • Soriano, Francesc X.;Baxter, Paul;Murray, Lyndsay M.;Sporn, Michael B.;Gillingwater, Thomas H.;Hardingham, Giles E.
    • Molecules and Cells
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    • v.27 no.3
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    • pp.279-282
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    • 2009
  • "Two-cysteine" peroxiredoxins are antioxidant enzymes that exert a cytoprotective effect in many models of oxidative stress. However, under highly oxidizing conditions they can be inactivated through hyperoxidation of their peroxidatic active site cysteine residue. Sulfiredoxin can reverse this hyperoxidation, thus reactivating peroxiredoxins. Here we review recent investigations that have shed further light on sulfiredoxin's role and regulation. Studies have revealed sulfiredoxin to be a dynamically regulated gene whose transcription is induced by a variety of signals and stimuli. Sulfiredoxin expression is regulated by the transcription factor AP-1, which mediates its up-regulation by synaptic activity in neurons, resulting in protection against oxidative stress. Furthermore, sulfiredoxin has been identified as a new member of the family of genes regulated by Nuclear factor erythroid 2-related factor (Nrf2) via a conserved cis-acting antioxidant response element (ARE). As such, sulfiredoxin is likely to contribute to the net antioxidative effect of small molecule activators of Nrf2. As discussed here, the proximal AP-1 site of the sulfiredoxin promoter is embedded within the ARE, as is common with Nrf2 target genes. Other recent studies have shown that sulfiredoxin induction via Nrf2 may form an important part of the protective response to oxidative stress in the lung, preventing peroxiredoxin hyperoxidation and, in certain cases, subsequent degradation. We illustrate here that sulfiredoxin can be rapidly induced in vivo by administration of CDDO-TFEA, a synthetic triterpenoid inducer of endogenous Nrf2, which may offer a way of reversing peroxiredoxin hyperoxidation in vivo following chronic or acute oxidative stress.

TI-I-174, a Synthetic Chalcone Derivative, Suppresses Nitric Oxide Production in Murine Macrophages via Heme Oxygenase-1 Induction and Inhibition of AP-1

  • Kim, Mi Jin;Kadayat, Taraman;Kim, Da Eun;Lee, Eung-Seok;Park, Pil-Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.390-399
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    • 2014
  • Chalcones (1,3-diaryl-2-propen-1-ones), a flavonoid subfamily, are widely known for their anti-inflammatory properties. Propenone moiety in chalcones is known to play an important role in generating biological responses by chalcones. In the present study, we synthesized chalcone derivatives structurally modified in propenone moiety and examined inhibitory effect on nitric oxide (NO) production and its potential mechanisms. Among the chalcone derivatives used for this study, TI-I-174 (3-(2-Hydroxyphenyl)-1-(thiophen-3-yl)prop-2-en-1-one) most potently inhibited lipopolysaccharide (LPS)-stimulated nitrite production in RAW 264.7 macrophages. TI-I-174 treatment also markedly inhibited inducible nitric oxide synthase (iNOS) expression. However, TI-I-174 did not significantly affect production of IL-6, cyclooxygenase-2 (COX-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that TI-I-174 inhibits production of inflammatory mediators in a selective manner. Treatment of macrophages with TI-I-174 significantly inhibited transcriptional activity of activator protein-1 (AP-1) as determined by luciferase reporter gene assay, whereas nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not affected by TI-I-1744. In addition, TI-I-174 significantly inhibited activation of c-Jun-N-Terminal kinase (JNK) without affecting ERK1/2 and p38MAPK, indicating that down-regulation of iNOS gene expression by TI-I-174 is mainly attributed by blockade of JNK/AP-1 activation. We also demonstrated that TI-I-174 treatment led to an increase in heme oxygenase-1 (HO-1) expression both at mRNA and protein level. Transfection of siRNA targeting HO-1 reversed TI-I-174-mediated inhibition of nitrite production. Taken together, these results indicate that TI-I-174 suppresses NO production in LPS-stimulated RAW 264.7 macrophages via induction of HO-1 and blockade of AP-1 activation.

Genetic Variation in Glutamate Carboxypeptidase II and Interaction with Dietary Natural Vitamin C May Predict Risk for Adenomatous Polyp Occurrence

  • Choi, Jeong-Hwa;Yates, Zoe;Martin, Charlotte;Boyd, Lyndell;Ng, Xiaowei;Skinner, Virginia;Wai, Ron;Kim, Jeongseon;Woo, Hae Dong;Veysey, Martin;Lucock, Mark
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4383-4386
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    • 2015
  • Background: The C1561T variant of the glutamate carboxypeptidase II (GCPII) gene is critical for natural methylfolylpolyglutamte (methylfolate) absorption, and has been associated with perturbations in folate metabolism and disease susceptibility. However, little is known on C1561T-GCPII as a risk factor for colorectal cancer. Therefore, this study examined whether C1561T-GCPII influences folate metabolism and adenomatous polyp occurrence. Materials and Methods: 164 controls and 38 adenomatous polyp cases were analysed to determine blood folate and plasma homocysteine (Hcy) level, dietary intake of natural methylfolate, synthetic pteroylglutamic acid (PteGlu), vitamin C and C1561T-GCPII genotype. Results: In controls and cases, 7.3 and 18.4 percent of subjects respectively, were found to have the CT genotype, increasing the risk for adenomatous polyp occurrence 2.86 times (95% CI:1.37-8.0, p=0.035). Total dietary folate, methylfolate and PteGlu intake and the level of erythrocyte folate and plasma Hcy did not predict the occurrence of an adenomatous polyp. However, dietary natural vitamin C intake was associated with adenomatous polyp risk within C1561T-GCPII CT genotype subjects (p=0.037). Conclusions: The findings suggest that C1561T-GCPII variation may be associated with risk for adenomatous polyp, and vitamin C may modify risk by interacting with the variant gene, its expression product and/or folate substrates.

Effects of spThoc7 Deletion on Growth and mRNA Export in Fission Yeast (분열효모에서 spThoc7 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.249-253
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    • 2014
  • THOC7/Mft1 is a member of the THO complex that is an evolutionally conserved assembly connecting transcription elongation with mRNP packaging and mRNA export. In fission yeast Schizosaccharomyces pombe, an ortholog (spThoc7) of THOC7/Mft1 was isolated by partial complementation of the lethality in synthetic lethal mutant, SLRsm1. A deletion mutant in a diploid strain was constructed by replacing one of spThoc7-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spthoc7 is nonessential for growth. But the ${\Delta}thoc7$ null mutant showed slight defects of both growth and mRNA export. And the functional spThoc7-GFP protein is localized mainly in the nucleus. These results suggest that spThoc7 is also involved in mRNA export from the nucleus to cytoplasm.

THI 52 Inhibits Inducible Nitric Oxide Synthase Gene Expression in RAW 264.7 Cells and Rat Lung Tissue by Lipopolysaccharide

  • Lee, Bog-Kyu;Park, Min-Kyu;Seo, Han-Geuk;YunChoi, Hye-Sook;Lee, Duck-Hyung;Chang, Ki-Churl
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.5
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    • pp.443-449
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    • 2001
  • Previously we reported that THI 52 inhibits tumor necrosis factor $(TNF)-{\alpha}$ mRNA expression in mouse peritoneal macrophages exposed to LPS plus $IFN-{\gamma}.$ In the present study, the effects of THI 52 on vascular reactivity ex vivo, and iNOS protein expression (rat lung) were investigated in LPS-treated rats. Treatment of THI 52 concentration-dependently reduced not only serum nitrite production but also the expression of iNOS protein in rat lung tissues. Thoracic aorta taken from LPS injected rat for 8 h ex vivo resulted in suppression of vasoconstrictor effects to phenylephrine (PE), which was restored by THI 52 (20 mg/kg) 30 min prior to LPS. When measured iNOS activity, treatment of THI 52 concentration-dependently reduced the enzyme activity in RAW 264.7 cells activated with LPS plus $IFN-{\gamma}.$ Likewise, iNOS activity was significantly reduced in lung tissues taken those rats that were injected THI 52 prior to LPS injection compared with LPS injection alone. These results strongly suggest that THI 52 can suppress iNOS gene expression induced by LPS, and restore the vascular contractility to PE. Thus, THI 52, a new synthetic isoquinoline alkaloid, may be beneficial in inflammatory disorders where production of NO is excessed by iNOS expression.

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Isolation and Characterization of a Theta Glutathione S-transferase Gene from Panax ginseng Meyer

  • Kim, Yu-Jin;Lee, Ok-Ran;Lee, Sung-Young;Kim, Kyung-Tack;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.449-460
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    • 2012
  • Plants have versatile detoxification systems to encounter the phytotoxicity of the wide range of natural and synthetic compounds present in the environment. Glutathione S-transferase (GST) is an enzyme that detoxifies natural and exogenous toxic compounds by conjugation with glutathione (GSH). Recently, several roles of GST giving stress tolerance in plants have demonstrated, but little is known about the role of ginseng GSTs. Therefore, this work aimed to provide further information on the GST gene present in Panax ginseng genome as well as its expression and function. A GST cDNA (PgGST) was isolated from P. ginseng cDNA library, and it showed the amino acid sequence similarity with theta type of GSTs. PgGST in ginseng plant was induced by exposure to metals, plant hormone, heavy metals, and high light irradiance. To improve the resistance against environmental stresses, full-length cDNA of PgGST was introduced into Nicotiana tabacum. Overexpression of PgGST led to twofold increase in GST-specific activity compared to the non-transgenic plants, and the GST overexpressed plant showed resistance against herbicide phosphinothricin. The results suggested that the PgGST isolated from ginseng might have a role in the protection mechanism against toxic materials such as heavy metals and herbicides.

A Visualization and Inference System for Protein-Protein Interaction (단백질 상호작용 추론 및 가시화 시스템)

  • Lee Mi-Kyung;Kim Ki-Bong
    • Journal of KIISE:Software and Applications
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    • v.31 no.12
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    • pp.1602-1610
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    • 2004
  • As various genome projects have produced enormous amount of biosequence data, functional sequence analysis in terms of tile nucleic acid and protein becomes very significant. In functional genomics and proteomics, the functional analysis of each individual gene and protein remains a big challenge. Contrary to traditional studies, which regard proteins as not components of a whole protein interaction network but individual entities, recent studies have focused on examining functions and roles of each individual gene and protein in view of a whole life system. In this regard, it has been recognized as an appropriate method to analyze protein function on the basis of synthetic information of its interaction and domain modularity. In this context, this paper introduces the PIVS (Protein-protein interaction Inference & Visualization System), which predicts the interaction relationship of input proteins by taking advantage of information on homology degree, domain modules which input sequences contain, and protein interaction relationship. The information on domain modules can increase the accuracy of the function and interaction relationship analysis in terms of the specificity and sensitivity.

Heterologous Expression and Characterization of a Laccase from Laccaria bicolor in Pichia pastoris and Arabidopsis thaliana

  • Wang, Bo;Yan, Ying;Xu, Jing;Fu, Xiaoyan;Han, Hongjuan;Gao, Jianjie;Li, Zhenjun;Wang, Lijuan;Tian, Yongsheng;Peng, Rihe;Yao, Quanhong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2057-2063
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    • 2018
  • Laccases can oxidize a variety of phenolic and non-phenolic substrates including synthetic dyes. In this research, a laccase gene Lcc9 from Laccaria bicolor was chemically synthesized and optimized to heterogeneous expression in Pichia pastoris and Arabidopsis thaliana. The properties of recombinant laccase expressed by P. pastoris were investigated. The laccase activity was optimal at 3.6 pH and $40^{\circ}C$. It exhibited $K_m$ and $V_{max}$ values of $0.565mmol\;l^{-1}$ and $1.51{\mu}mol\;l^{-1}\;min^{-1}$ for ABTS respectively. As compared with untransformed control plants, the laccase activity in crude extracts of transgenic lines exhibited a 5.4 to 12.4-fold increase. Both laccases expressed in transgenic P. pastoris or A. thaliana could decolorize crystal violet. These results indicated that L. bicolor laccase gene may be transgenically exploited in fungi or plants for dye decolorization.