• Title/Summary/Keyword: Tissue-specific expression

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Functional properties of an alternative, tissue-specific promoter for rice NADPH-dependent dihydroflavonol reductase

  • Kim, Joonki;Lee, Hye-Jung;Tyagi, Wricha;Kovach, Michael;Sweeney, Megan;McCouch, Susan;Cho, Yong-Gu
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.163-163
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    • 2017
  • A deletion analysis of the Oryza sativa dihydroflavonol reductase (DFR) promoter defined a 25 bp region (-386 to -362) sufficient to confer pericarp-specific expression of ${\beta}$ -glucuronidase(GUS) reporter gene in transgenic rice. Site-specific mutagenesis of these conserved sequences and subsequent expression analysis in calli which transiently expressed the mutated promoter::GUS gene showed that both bHLH (-386 to -381) and Myb (-368 to -362) binding sites in the DEL3 (-440 to 70) promoter were necessary for complete expression of the GUS gene including the tissue-specific expression of DFR::GUS gene. The GUS gene was expressed well in the mutated Myb (-368 to -362) binding site, but not as strong as in normal condition, implying that the Myb is also necessary to express GUS gene fully. Also, we found the non-epistatic relation between Rc and DFR. There were no changes of expression patterns GUS under the Rc and rc genotypes. Thus, DFR expression might be independent of the presence of functional Rc gene and suggested that Rc and Rd (DFR) share the same pathway controlling the regulation of flavonoid synthesis but not a direct positive transcriptional regulator of DFR gene.

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Regulatory Mechanism in Tissue-specific Expression of Insulin-like Growth Factor-I Gene (Insulin-like growth factor-I 유전자의 조직 특이적 발현에 대한 조절기전)

  • 안미라
    • KSBB Journal
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    • v.18 no.4
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    • pp.329-334
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    • 2003
  • The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBP${\alpha}$ and C/EBP${\beta}$ proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\alpha}$/, p35$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\beta}$/, and p35$\^$C/EBP${\beta}$ form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42$\^$C/EBP${\alpha}$/ and p38$\^$C/EBP${\beta}$/ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.

Tissue- and Reproductive Organ-specific Expression of Protease Nexin-1 in Sprague-Dawley Rat (흰쥐에서 단백질 분해효소 저해제, Nexin-1의 조직 및 생식기관 특이적 유전자 발현)

  • 고정재;김남근;김진규;최명진;정형민;서승염;김윤희;이현환;차광열
    • Development and Reproduction
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    • v.2 no.2
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    • pp.135-140
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    • 1998
  • Protease Nexin-1 (PN-1) inhibits the activity of several serine proteases including thrombin, urokinase (uPA)-type plasminogen activator and trypsin. Tissue- and reproductive organ-specific mRNA levels of the PN-1 were investigated in Sprague-Dawley adult rat. PN-1 mRNA expression in rats was found in brain (forebrain, hindbrain), heart, liver, lung, ovary and oviduct. The level of PN-1 mRNA in male and female among the tissues was the highest in forebrain of the female. PN-1 expression in reproductive organs was found only in ovary and oviduct. These results suggest that PN-1 expression is dependent on the sex and may be related to folliculogenesis and early embryogenesis.

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Modulation of TNFSF expression in lymphoid tissue inducer cells by dendritic cells activated with Toll-like receptor ligands

  • Han, Sin-Suk;Koo, Ji-Hye;Bae, Jin-Gyu;Kim, Soo-Chan;Baik, Song;Kim, Mi-Yeon
    • BMB Reports
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    • v.44 no.2
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    • pp.129-134
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    • 2011
  • Toll-like receptors (TLRs), which recognize structurally conserved components among pathogens, are mainly expressed by antigen-presenting cells such as dendritic cells (DCs), B cells, and macrophages. Recognition through TLRs triggers innate immune responses and influences antigen-specific adaptive immune responses. Although studies on the expression and functions of TLRs in antigen-presenting cells have been extensively reported, studies in lymphoid tissue inducer (LTi) cells have been limited. In this study, we observed that LTi cells expressed TLR2 and TLR4 mRNA as well as TLR2 protein and upregulated OX40L, CD30L, and TRANCE expression after stimulation with the TLR2 ligand zymosan or TLR4 ligand LPS. The expression of tumor necrosis factor superfamily (TNFSF) members was significantly upregulated when cells were cocultured with DCs, suggesting that upregulated TNFSF expression may contribute to antigen-specific adaptive immune responses.

Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle

  • Baik, M.;Vu, T.T.T.;Piao, M.Y.;Kang, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1493-1498
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    • 2014
  • Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

Expression of collagenases (matrix metalloproteinase-1, 8, 13) and tissue inhibitor of metalloproteinase-1 of retrodiscal tissue in temporomandibular joint disorder patients

  • Gho, Won Gyung;Choi, Yuri;Park, Kwang-Ho;Huh, Jong-Ki
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.44 no.3
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    • pp.120-127
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    • 2018
  • Objectives: The aim of this study was to reveal how collagenases (matrix metalloproteinase [MMP]-1, 8, 13) and tissue inhibitor of metalloproteinase 1 (TIMP-1) are expressed in immunohistochemistry of retrodiscal tissue in temporomandibular joint disorder patients. Materials and Methods: This study was conducted on 39 patients who underwent discoplasty or discectomy. Immunohistochemical staining was undertaken and expression levels of MMP-1, 8, 13, and TIMP-1 were evaluated. The status of internal derangement of disc, osteoarthritis, and joint effusion were analyzed using magnetic resonance imaging (MRI). Disc status observed during operation was also categorized. Results: The more severe disc derangement was observed on MRI, the more increased expression of MMPs and TIMP-1 appeared. Regarding MMP-13 expression, 86.7% of late-stage disc displacement patients showed grade II or III. Expression level of MMPs or TIMP was not statistically significant associated with joint effusion level. In perforation and/or adhesion groups, all patients showed grade II or III expression of MMP-13. Once perforation occurred, MMP-13 showed increased expression with statistical significance. Conclusion: MMP-1 and MMP-13 expression seem to be related to progression of osteoarthritis whereas MMP-8 does not seem to have a specific role with regard to temporomandibular joint disorders. TIMP-1 is considered to be partly related to internal derangement rather than osteoarthritis, but it is not significant.

Hemocyte-specific Promoter for the Development of Transgenic Silkworm, Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Choi, Gwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.1
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    • pp.111-114
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    • 2012
  • In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.

Expression of mRNA for matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gingival and periodontal ligament fibroblasts treated with lipopolysaccharide from Prevotella intermedia (Prevotella intermedia의 세균내독소가 치은섬유아세포와 치주인대세포에서의 matrix metalloproteinase 및 tissue inhibitor of metalloproteinase의 발현에 미치는 영향)

  • Kim, Sung-Jo;Choi, Eun-Young;Choi, In-Soon;Lee, Ju-Youn;Choi, Jeom-Il;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.35 no.1
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    • pp.21-30
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    • 2005
  • Matrix metalloproteinases (MMPs) are a family of host-derived proteolytic enzymes and implicated in the remodeling and degradation of extracellular matrix under both physiological and pathological conditions. Connective tissue degradation in periodontal diseases is thought to be due to excessive MMP activities over their specific inhibitors. The effects of lipopolysaccharide (LPS) from Prevotella intermedia, one of the major putative pathogens of periodontitis, on the expression of mRNA for MMPs and tissue inhibitors of metalloproteinases (TIMPs) in human gingival and periodontal ligament fibroblasts were examined by reverse transcriptase-polymerase chain reaction (RT-PCR). The expression of mRNAs encoding MMP-1, -2, -3, -10, and -14 was increased in human gingival fibroblasts treated with p. intermedia LPS, whereas MMP-11 and TIMP-2 mRNA expression was decreased in these cells stimulated with LPS. P. intermedia LPS increased the MMP-1, -2, -10, -11, and -14 mRNA expression and decreased TIMP-1 and -2 mRNA expression in human periodontal ligament fibroblasts. These findings imply that P. intermedia LPS may play an important role in the connective tissue degradation in periodontitis.

Development of Transgenic Fish for the Production of Human EGF Protein (내재적 유전자에 의한 어류난자에서의 hEGE 단백질 생산을 위한 기술개발)

  • 황창남;송기철;이재현;윤종만;김기동;이상호;박홍양
    • Korean Journal of Animal Reproduction
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    • v.25 no.3
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    • pp.277-286
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    • 2001
  • Improvement and possible commercialization of a home-made electroporation apparatus(home-made) were further tried to establish a simple and effective introduction of foreign gene into sperm followed by in vitro fertilization. Expressions of introduced pJJ9 and pNT plasmids were shown in all fertilized eggs with electroporated spermatozoa. In particular, with this gene transfer system all the fry showed a consistently transient expression in the syncytium of the yolk sac. This fact is important since some required, minute quantity of human proteins can be produced from the established transient expression on the yolk sac of all fry derived from in vitro fertilization with electroporated spermatozoa. To explore tissue-specific expression in fish, which we will use a similar system later, we targeted the nerve tissue to see whether tissue-specific promoter is working in fish properly. pNT plasmid containing a nerve cell-specific tubulin promoter gene demonstrated consistently exact targeted expressions among the developing nerve cells in later stages of embryos and hatched fry. Finally, liver-specific genes are now being cloned by using already selected primers for useful human protein gene fusion.

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Peroxisome Proliferator-activated Receptor ${\gamma}$ Is Not Associated with Adipogenesis in Female Mice

  • Yoon, Mi-Chung;Jeong, Sun-Hyo
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.139-146
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    • 2008
  • The peroxisome proliferator-activated receptor ${\gamma}$ $(PPAR{\gamma})$ plays a central role in adipogenesis and lipid storage. The $(PPAR{\gamma})$ ligands, thiazolidinediones (TZDs), enhance in vitro adipogenesis in several cell types, but the role of the TZDs on in vivo adipogenesis is still poorly understood. To investigate how $PPAR{\gamma}$ ligand troglitazone regulates adipogenesis in female mice, we examined the effects of the troglitazone on adipose tissue mass, morphological changes of adipocytes, and the expression of $PPAR{\gamma}$ target and adipocyte-specific genes in low fat diet-fed female C57BL/6 mice. Administration of troglitazone for 13 weeks did not change body and total white adipose tissue weights compared with control mice. Troglitazone treatment also did not cause a significant decrease in the average size of adipocytes in parametrial adipose tissue although it is reported to increase the number of small adipocytes in male animals. Troglitazone did not affect the mRNA expression of $PPAR{\gamma}$ and its target genes as well as adipocyte-specific genes in parametrial adipose tissue. These results suggest that $PPAR{\gamma}$ does not seem to be associated with adipogenesis in females with functioning ovaries and that its inability to induce adipogenesis may be due to sex-related factors.

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