• Title/Summary/Keyword: Nuclear localization signal

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Structure and Tissue Distribution of a Trinucleotide-Repeat-containing Gene (cag-3) Expressed Specifically in the Mouse Brain

  • Ji, Jin Woo;Yang, Hye Lim;Kim, Sun Jung
    • Molecules and Cells
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    • v.20 no.3
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    • pp.348-353
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    • 2005
  • Using in silico approaches and RACE we cloned a full length trinucleotide (CAG) repeat-containing cDNA (cag-3). The cDNA is 2478 bp long and the deduced polypeptide consists of 140 amino acids of which 73 are glutamines. The genomic sequence spans approximately 79 kb on mouse chromosome 7 and the gene is composed of four exons. Standard and real-time PCR analyses of several mouse tissues showed that the gene is exclusively expressed in the brain and is not detected in embryonic stages. Within the brain, it is expressed throughout the forebrain region with predominant expression in the hypothalamus and olfactory bulb and very low levels in the mid- and hindbrain.

Shielded High-Order Gradient Coil Design for Magnetic Resonance Spectroscopy and Imaging (자기공명분석과 영상촬영을 위한 차폐된 고차경사자계코일의 설계)

  • Oh, Chang-Hyun;Hilal, Sadek K.;Yi, Yun;Kim, Min-Gi
    • Proceedings of the KOSOMBE Conference
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    • v.1993 no.05
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    • pp.45-48
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    • 1993
  • High-order field gradients are useful for spatial localization of a volume of interest and dynamic range improvement of signal detection in NMR (Nuclear Magnetic Resonance) spectroscopy and imaging. This paper proposes a design method of shielded high-order gradient coils to reduce tile effect of eddy current on tile spectroscopy and imaging results. According to the experimental results, the shielded gradient coils produce less than 2 % eddy current compared to non-shielded coils. Two shielded $z^2$ gradient coils have been designed and constructed for 1.5 T whole-body and 3.0 T animal NMR imaging systems. Experimental results are in good agreement with the theoretically expected behavior and show the utility of the shielded high-order gradient coils.

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The innate immune response transcription factor Bombyx mori Relish1 induces high-level antimicrobial peptides in silkworm

  • Kim, Seong-Wan;Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.37 no.2
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    • pp.49-54
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    • 2018
  • To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.

A DPL (DNA/peptide/liposomes) Tripartite Complex Effective for Transfection in Serum

  • Kim Young-Cheol;Park Jong-Gu
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.187-194
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    • 2004
  • A short peptide corresponding to the nuclear localization signal (NLS) of human immunodeficiency virus (HIV)-l Tat protein, Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg, was employed to improve the efficiency of cellular uptake of nucleic acids. The peptide was first mixed with a reporter plasmid and then with cationic liposomes to form a tripartite complex of DNA/peptide/liposomes (DPL). Transfection efficiency of the DPL complex was compared with that of the conventional DNA/liposomes (DL) complex. When the DPL complex was formed with various cationic liposomes, DOTAP/DOPE (DP) liposome exhibited superior transfection efficiency to other liposomes tested in vitro. With the inclusion of the peptide, the DPL complex showed much enhanced transfection in various cancer cell lines. Particularly, transfection of the DPL complex in serum increased cellular uptake of a transgene up to 2 fold when compared with that in a serum free condition. Further, when the DPL complex was infused through the ureteric route of a rat, transfection efficiency was shown to be better in reporter gene expression than that obtained with the DL complex. This study shows that the DPL complex that is easy to formulate can be employed for much enhanced cellular uptake of a trans gene.

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Cloning and Expression Analysis of Gonadogenesis-associated Gene SPATA4 from Rainbow Trout (Oncorhynchus mykiss)

  • Liu, Bowen;Liu, Shangfeng;He, Shan;Zhao, Ying;Hu, Hongxia;Wang, Zhao
    • BMB Reports
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    • v.38 no.2
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    • pp.206-210
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    • 2005
  • Gonadogenesis is a complicated process which involves multi-gene interactions. A rainbow trout (Oncorhynchus mykiss) gene spermatogenesis associated 4 (SPATA4) was cloned and characterized from adult rainbow trout testis. The cDNA sequence of rainbow trout SPATA4 contains an open reading frame of 1, 081 nucleatides encoding a putative protein of 259 amino acids. The putative protein from rainbow trout shares a 76.8% homology with zebrafish SPATA4. No trans-membrane regions or signal peptide were detected using bioinformatics methods. Subcellular localization analysis revealed that rainbow trout SPATA4 was a nuclear protein with highest possibility (39.1%). Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the distribution of rainbow trout SPATA4 in eleven organs of adult rainbow trout. The result demonstrated that this gene express specifically in testis and slight amount of expression was detected in ovary. Further analysis of SPATA4 characterization and function in rainbow trout may provide insight into the understanding of gonadogenesis process.

Performances of the Directional Control Solenoid Valve for a Combined Power Plant

  • Kim, Chul-Jin;Yun, Yu-Seong;Kim, Do-Tae;Lee, Il-Young
    • International Journal of Safety
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    • v.11 no.2
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    • pp.10-14
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    • 2012
  • Recently, the combined power plants are refocused rapidly as a replaceable energy system of the nuclear power plant. The large turbine is revolved highly at 1800~3600 rpm. Thus, the turbine speed should be monitored with mechanical and electrical method for a safety. The electrical cutoff valve which blocks the flow channel with the electrical signal is with a built in. The aim of this study is to develop a manufacturing technology through by the localization of a solenoid actuated directional control valve. Especially the results show performances of the solenoid valve by the experiments and modeling and the reliability estimation. Applied load port pressure was changed rapidly on the form of a quadratic curve over time. And in the cases of square waveform when 0~100 V and 20~120 V input voltage, it was driven on a stable state until 13.4 Hz and 16.6 Hz, respectively. We think that this study will give useful data for the electricity safety system of the combined power plant gas turbine.

Molecular Cloning and Characterization of a Heat Shock Protein 70-related cDNA from Olive flounder (Paralichthys olivaceus)

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Lee, Sang-Jun;Kang, Ho-Sung;Kim, Han-Do
    • Journal of Aquaculture
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    • v.12 no.2
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    • pp.91-100
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    • 1999
  • The complete nucleotide sequence of olive flounder (Paralichthys olivaceus) hsp70-related rDNA was determined by RT- and RACE-PCR methods. A full-length of hsp70-related cDNA has an open reading frame of 1.95 kb encoding 650 amino acids with a calculated molecular weight of 71.1 kD. A corresponding hsp70-related protein contains a number of conserved elements including an ATP-binding domain, a nuclear localization signal and the carboxyl terminal motif, EEVD, which may have a role in chaperone function. Comparison of nucleotide and predicted amino acid sequence between olive flounder hsp70-related gene and hsp/hsc70 genes of other species revealed a high similarity with the cognate form of these genes. These results indicated that we recovered likely to be a olive flounder cognate hsc70 gene.

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Ectopic Overexpression of Coiled-Coil Domain Containing 110 Delays G2/M Entry in U2-OS Cells

  • Lee, Sue Nyoung;Hong, Kyeong-Man;Seong, Yeon Sun;Kwak, Sahng-June
    • Development and Reproduction
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    • v.24 no.2
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    • pp.101-111
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    • 2020
  • Coiled-coil domain containing 110 (CCDC110, KM-HN-1) is a protein containing C-terminal coiled-coil domain (CCD) which was previously discovered as a member of the human cancer/testis antigen (CTA). In addition, CCDC110 has both nuclear localization signal sequence and the leucine zipper motif. Although the functional role of CCDC110 has yet to be fully identified, the mRNA expression levels of CCDC110 are known to be highly elevated in various cancer types including testis, implying its relevance to cancer pathogenesis. In this study, we first developed several monoclonal antibody (mAb) hybridoma clones targeting CCDC110 and further isolated clone by characterizing for its specificity using immunoblotting and immunoprecipitation approaches with basal parenchymal sperm cells in testis tissue. Next, using these mAbs, we showed that the Tet-inducible overexpression of CCDC110 protein delayed the entry of G2/M phase in U2-OS osteosarcoma cells. Based on these results, we propose that CCDC110 plays a crucial role in cell cycle progression.

A Basic Helix-Loop-Helix Transcription Factor Regulates Cell Elongation and Seed Germination

  • Kim, Jin-A;Yun, Ju;Lee, Minsun;Kim, Youn-Sung;Woo, Jae-Chang;Park, Chung-Mo
    • Molecules and Cells
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    • v.19 no.3
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    • pp.334-341
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    • 2005
  • Plants are sessile and rely on a wide variety of growth hormones to adjust growth and development in response to internal and external stimuli. We have identified a gene, designated NAN, encoding a basic helix-loop-helix (bHLH) transcription factor that regulates cell elongation and seed germination in plants. NAN has an HLH motif in its C-terminal region but does not have any other discernible homologies to bHLH proteins. A bipartite nuclear localization signal is located close to the HLH motif. An Arabidopsis mutant, nan-1D, in which NAN is activated by the insertion of the 35S enhancer, exhibits growth retardation with short hypocotyls and curled leaves. It is also characterized by reduced seed germination and apical hook formation, symptomatic of GA deficiency or disrupted GA signaling. The phenotypic effects of nan-1D were increased by treatment with paclobutrazol (PAC), an inhibitor of gibberellic acid (GA) biosynthesis. NAN is constitutively expressed throughout the life cycle. Our observations indicate that NAN has a housekeeping role in plant growth and development, particularly in seed germination and cell elongation, and that it may modulate GA signaling.