• Title/Summary/Keyword: Light chain gene

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Comparative Study of Gene Expression Profiles in Posterior Silk Glands of the Silkworm, Bombyx mori L.

  • Choi, Kwang-Ho;Goo, Tae-Won;Kang, Seok-Woo;Kang, Min-Uk;Yun, Eun-Young;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.229-234
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    • 2008
  • We used serial analysis of gene expression (SAGE) approach to derive a profile of expressed genes of the posterior silk glands (PSG) and to create a reference for understanding gene cluster related to the mechanism of silk protein synthesis in the silkworm, Bombyx mori. We constructed a 3' SAGE library from the PSG of the fifth instar larvae of the silkworm. In total we obtained 2,406 SAGE tags, of which 682 were unique tags. Sorted by tag count number, 27 (4%) unique tags were significantly more abundant genes (ten or more times), whereas 445 (65%) unique tags were detected as single copies. The annotation of 682 unique SAGE tags revealed that 462 (68%) of the SAGE tag sequences represented known genes, whereas 220 (32%) of the tag sequences had no matches in SAGE map and silkworm EST databases. Of the 682 SAGE tags, the most abundant tag sequences were that of the fibroin light chain gene and the silk protein P25. In addition, we compared two relative abundance results of the SAGE and the EST approaches to verify whether their transcript quantitative aspects are significant or not. The comparative results of relative abundances of the fibroin H-, L- chain and P25 glycoprotein genes indicated that the quantitative approach based on SAGE tags is effective for quantitative cataloging and comparison of expressed genes in same organs. The SAGE tag information reported in this study would be useful for researchers in the field to analyze genes associated with silk processing mechanisms of insects.

Agronomic characteristics of stay-green mutant derived from an early-maturing rice variety 'Pyeongwon'

  • Won, Yong-Jae;Ji, Hyeon-So;Ahn, Eok-Keun;Lee, Jeong-Heui;Jung, Kuk-Hyun;Lee, Sang-Bok;Hong, Ha-Cheol;Hyun, Ung-Jo;Ha, Woon-Goo;Kim, Myeong-Ki;Kim, Byeong-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.72-72
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    • 2017
  • We found a new stay-green mutant from 'Pyeongwon' which is an early-maturing rice variety in Korea. The mutant showed green leaves after grain ripening period and it maintained higher SPAD value than wild type rice plant and original variety 'Pyeongwon'. The stay-green trait in rice, three genes have been identified up to date. The non-yellow coloring1 (NYC1) gene encodes a chloroplast-localized short-chain dehydrogenase/reductase (SDR) with three transmembrane domains. The non-yellow coloring3 (NYC3) gene encodes a plastid-localizing alpha/beta hydrolase-fold family protein with an esterase/lipase motif. The Sgr gene encodes a novel chloroplast protein and regulates the destabilization of the light-harvesting chlorophyll binding protein (LHCP) complexes of the thylakoid membranes, which is a prerequisite event for the degradation of chlorophylls and LHCPs during senescence. After sequencing the PCR products, we found a single nucleotide variation($A{\rightarrow}T$) in the NYC1 gene, which changes the amino acid lysine to methionine. The NYC1 gene encodes a short-chain dehydrogenase/reductase(SDR) protein. And we confirmed the co-segregation between SNP and stay-green trait from genotyping the progenies of the mutant.

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Effects of 630-nm Organic Light-emitting Diodes on Antioxidant Regulation and Aging-related Gene Expression Compared to Light-emitting Diodes of the Same Wavelength

  • Mo, SangJoon;Kim, Eun Young;Ahn, Jin Chul
    • Current Optics and Photonics
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    • v.6 no.3
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    • pp.227-235
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    • 2022
  • To investigate the aging-related physiological functions of organic light-emitting diodes (OLEDs), we examined mRNA expression changes in aging-related genes due to oxidative stress inhibition by 630-nm red light OLEDs. As a result of irradiating 630-nm OLED with an intensity of 5 mW/cm2 for 15 min, the viability of dermal fibroblasts significantly increased by 1.3-fold. In addition, reactive oxygen species generated by H2O2 were significantly reduced about 4.9-fold by irradiation with 630-nm OLED. Quantitative reverse-transcription polymerase chain reaction results showed that 630-nm OLEDs altered aging-related gene mRNA expression levels through antioxidant activity. The mRNA expression levels of matrix metalloproteinase1 (MMP1) and MMP9 decreased significantly, by about 2.2- and 2.5-fold, compared to the control group, whereas those of collagen, type I, and alpha 1 increased significantly, by 4.9-fold. The mRNA expression levels of cancer suppression genes p16 and p53 in dermal fibroblasts were also significantly reduced by 630-nm OLED irradiation, by about 1.4- and three-fold, respectively, compared to the control. Overall, it was confirmed that 630-nm OLED irradiation lowered the level of ROS formation induced by H2O2 in dermal fibroblasts, and that this antioxidant effect could regulate the mRNA expression levels of aging- and tumor suppression-related genes. This study shows a link between 630-nm OLED irradiation and anti-aging physiological functions such as antioxidant function, and suggests the potential of OLEDs as a useful light source for skin care.

The Development of Chicken Recombinant Single-chain Fv (ScFv) Antibody Reactive with Sporozoite Antigen of Eimeria spp. which Causes Avian Coccidiosis (가금 콕시듐증을 일으키는 Eimeria spp.의 포자충 항원에 결합하는 닭의 재조합 항체(ScFv)의 개발)

  • Park, Dong-Woon;Kim, Eon-Dong;Kim, Sung-Heon;Han, Jae-Yong;Kim, Jin-Kyoo
    • Korean Journal of Poultry Science
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    • v.38 no.4
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    • pp.323-330
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    • 2011
  • The chicken monoclonal antibody (mAb), 13C8, reacts with sporozoite antigens of Eimeria spp. which causes avian coccidiosis. Since this mAb was produced at low amount due to genetic instability of chicken hybridoma, a recombinant 13C8 single-chain Fv (ScFv) antibody was constructed by amplification of the variable domain of heavy (VH) and light chain (VL) genes of antibody derived from chicken hybridoma. The constructed 13C8 ScFv was successfully expressed in E. coli and purified as a soluble form. In ELISA analysis, this recombinant 13C8 ScFv antibody showed antigen binding activity as the original mAb. In addition, nucleotide sequence comparison of 13C8 gene to the germline chicken VL and VH genes suggested that the gene conversion with $V{\lambda}$ and VH pseudogenes might contribute to the diversification of VL and VH genes in chickens.

The Rat Myosin Light Chain Promoter-Driven DsRed Reporter System Allows Specific Monitoring of Bone Marrow Mesenchymal Stem Cell- Derived Cardiomyocytes

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.21-25
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    • 2008
  • Bone marrow mesenchymal stem cells (BMMSCs) have the capacity for self-renewal and differentiation into a variety of cell types. They represent an attractive source of cells for gene and cell therapy. The purpose of this study is to direct the specific expression of the DsRed reporter gene in $Sca-1^+$ BMMSCs differentiated into a cardiomyogenic lineage. We constructed the prMLC-2v-DsRed vector expressing DsRed under the control of the 309 tp fragment of the rat MLC-2v 5'-flanking region. The specific expression of the DsRed reporter gene under the transcriptional control of the 309 bp fragment of the rat MLC-2v promoter was tested in 5-azacytidine healed-$Sca-1^+$ BMMSCs over 2 weeks after the prMLC-2v-DsRed transfection. The prMLC-2v-DsRed was specifically expressed in the $Sca-1^+$ BMMSCs with cardiomyogenic lineage differentiation and it demonstrates that the 309 bp sequences of the rat MLC-2v 5'-flanking region is sufficient to confer cardiac specific expression on a DsRed reporter gene. The cardiac-specific promoter-driven reporter vector provides an important tool for the study of stem cell differentiation and cell replacement therapy in ischemic cardiomyopathy.

Altered Gene Expression in Cerulein-Stimulated Pancreatic Acinar Cells: Pathologic Mechanism of Acute Pancreatitis

  • Yu, Ji-Hoon;Lim, Joo-Weon;Kim, Hye-Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.409-416
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    • 2009
  • Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with $10^{-8}$ M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.

Improved Coexpression and Multiassembly Properties of Recombinant Human Ferritin Subunits in Escherichia coli

  • Lee, Jung-Lim;Levin, Robert E.;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.926-932
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    • 2008
  • Human heavy chain (H-) and light chain (L-) ferritins were amplified from a human cDNA library. Each ferritin gene was inserted downstream of the T7 promoter of bacterial expression vectors, and two types of coexpression vectors were constructed. The expression levels of recombinant ferritins ranged about 26-36% of whole-cell protein. H-ferritin exhibited a lower expression ratio compared with L-ferritin, by a coexpression system. However, the coexpression of HL-ferritins was significantly increased above the expression ratio of H-ferritin by cultivation without IPTG induction overnight. Purified recombinant H-, L-, HL-, and LH-ferritins were shown to be homo- and heteropolymeric high molecular complexes and it was indicated that their assembled subunits would be able to work functionally in the cell. Thus, these results indicate an improvement in the expression strategy of H-ferritin for heteropolymeric production and studies of ferritin assembly in Escherichia coli.

Biochemical and molecular characterization of a tetrachloroethylene (PCE) dechlorinating Clostridium bifermentans DPH-1

  • Chang, Young-Cheol;Toyama, Tadashi;Kikuchi, Shintaro
    • Journal of environmental and Sanitary engineering
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    • v.23 no.2
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    • pp.1-18
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    • 2008
  • The tetrachloroethylene (PCE) dehalogenase of Clostridium bifermentans DPH-1 (a halorespiring organism) was purified, cloned, and sequenced. This enzyme is a homodimer with a molecular mass of ca. 70 kDa and exhibits dehalogenation of dichloroethylene isomers along with PCE and trichloroethylene (TCE). Broad range of substrate specificity for chlorinated aliphatic compounds (PCE, TCE, cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,2-dichloropropene, and 1,1,2-trichloroethane) for this enzyme was also observed. A mixture of propyl iodide and titanium citrate caused a light-reversible inhibition of enzymatic activity suggesting the involvement of a corrinoid cofactor. A partial sequence (81 bp) of the encoding gene for PCE dehalogenase was amplified and sequenced with degenerateprimers designed from the N-terminal sequence (27 amino acid residues). Southern analysis of C. bifermentans genomic DNA using the polymerase chain reaction product as a probe revealed restriction fragment bands. A 5.0 kb ClaI fragment, harboring the relevant gene (designated pceC) was cloned (pDEHAL5) and the complete nucleotide sequence of pceC was determined. The gene showed homology mainly with microbial membrane proteins and no homology with any known dehalogenase, suggesting a distinct PCE dehalogenase. So, C. bifermentans could play some important role in the initial breakdown of PCE and other chlorinated aliphatic compounds in sites contaminated with mixtures of halogenated substances.

PHOTOMORPHOGENIC MUTANTS OF TOMATO

  • Kendrick, Richard E.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1994.09a
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    • pp.41-51
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    • 1994
  • Tomato (Lycopersicon esculentum Mill.) has been chosen as a model species for the study of hotomorphogenesis. The aurea (au) and yellow-green-2 (yg-2) mutants which are severely photochrome deficient appear to be phytochrome chromophore mutants. Mutants modified with respect to specific members of the phytochrome gene family: the far-red light-insensitive mutant (fri, for phytochrome A) and the temporarily red light-insensitive mutant (tri, for phytochrome B1) have been identified. Mutants that exhibit an exaggerated phytochrome response are putative transduction-chain mutants affecting an amplification step in phytochrome signal transduction. These mutants are being used to understand the complexities of juvenile anthocyanin in the hypocotyl during seedling de-etiolation.

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A Comparative Study of Gene Expression Patterns of Periodontal Ligament Cells and Gingival Fibroblasts using the cDNA Microarray (cDNA Microarray를 이용한 치주인대세포와 치은섬유아세포의 유전자 발현에 대한 연구)

  • Jeon, Chai-Young;Park, Jin-Woo;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.205-221
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    • 2004
  • Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.