• Title/Summary/Keyword: knockout

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Role of $\mu$-Opioid Receptors on Neurobehaviors (뇌신경행동에 미치는 $\mu$-opioid 수용체의 역할)

  • Jang, Choon-Gon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.04a
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    • pp.21-37
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    • 2003
  • 1. Stimulation of dopaminergic system by morphine was abolished in ${\mu}$-opioid receptor knockout mice. 2. Dopaminergic stimulation by opioid agonists, morphine, DPDPE, and U50488, acts independently. 3. Loss of ${\mu}$-opioid receptors is more sensitive to the response of NMDA-induced convulsion and increase in the expression of mRNA for NMDA receptors.

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Association of a Common Reductase with Multiple Aromatic Terminal Dioxygenases in Sphingomonas yanoikuyae Strain B1

  • Mihyun Bae;Kim, Eungbin
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.40-43
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    • 2000
  • The aromatic dioxygenase system in Sphingomonas yanoikuyae strain Bl consists of three components, an oxygenase, a ferredoxin, and a reductase. The insertional knockout of the bphA4 gene encoding a reductase and subsequent complementation experiments showed that the reductase encoded by bphA4 in S. yanoikuyae strain Bl is associated with multiple dioxygenase components including that of toluate dioxygenase (XyIXY).

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Effect of Several Endocrine Disrupting Compound on Mammary Gland Carcinogenesis in c-Ha-ras-trasgenic Rats

  • Han, Bum-Sup
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2001.09a
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    • pp.13-15
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    • 2001
  • 발암성시험연구에 사용되고 있는 형질전환 동물들은 랫드와 마우스 등이 있는데, 그 중 c-Ha-ras proto-oncogene 마우스 (ras H2 mice), v-Ha-ras 형질전환 마우스 (Tg.AC mice), pim-1 형질전환 마우스 및 p53 knockout 마우스 등이 발암유발물질에 감수성이 높아 현재 중기발암성시험에 이용되고 있다. (중략)

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Differential Subcellular Localization of Ribosomal Protein L7 Paralogs in Saccharomyces cerevisiae

  • Kim, Tae-Youl;Ha, Cheol Woong;Huh, Won-Ki
    • Molecules and Cells
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    • v.27 no.5
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    • pp.539-546
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    • 2009
  • In Saccharomyces cerevisiae, ribosomal protein L7, one of the ~46 ribosomal proteins of the 60S subunit, is encoded by paralogous RPL7A and RPL7B genes. The amino acid sequence identity between RPl7a and RPl7b is 97 percent; they differ by only 5 amino acid residues. Interestingly, despite the high sequence homology, Rpl7b is detected in both the cytoplasm and the nucleolus, whereas Rpl7a is detected exclusively in the cytoplasm. A site-directed mutagenesis experiment revealed that the change in the amino acid sequence of Rpl7b does not influence its subcellular localization. In addition, introns of RPL7A and RPL7B did not affect the subcellular localization of Rpl7a and Rpl7b. Remarkably, Rpl7b was detected exclusively in the cytoplasm in rpl7a knockout mutant, and overexpression of Rpl7a resulted in its accumulation in the nucleolus, indicating that the subcellular localization of Rpl7a and Rpl7b is influenced by the intracellular level of Rpl7a. Rpl7b showed a wide range of localization patterns, from exclusively cytoplasmic to exclusively nucleolar, in knockout mutants for some rRNA-processing factors, nuclear pore proteins, and large ribosomal subunit assembly factors. Rpl7a, however, was detected exclusively in the cytoplasm in these mutants. Taken together, these results suggest that although Rpl7a and Rpl7b are paralogous and functionally replaceable with each other, their precise physiological roles may not be identical.

Differential Expression of Genes Important to Efferent Ductules Ion Homeostasis across Postnatal Development in Estrogen Receptor-α Knockout and Wildtype Mice

  • Lee, Ki-Ho;Bunick, David;Lamprecht, Georg;Choi, Inho;Bahr, Janice M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.4
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    • pp.510-522
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    • 2008
  • Our earlier studies showed that estrogen was involved in the regulation of fluid reabsorption in adult mouse efferent ductules (ED), through estrogen receptor (ER) ${\alpha}$ and $ER{\beta}$ by modulating gene expression of epithelial genes involved in ion homeostasis. However, little is known about the importance of $ER{\alpha}$ in the ED during postnatal development. Based on previous findings, we hypothesized that there should be a difference in the expression of epithelial ion transporters and anion producers in the ED of postnatal wild type (WT) and estrogen receptor ${\alpha}$ knockout (${\alpha}ERKO$) mice. Using absolute, comparative and semi-quantitative RT-PCR along with immunohistochemistry, we looked at expression levels of several genes in the ED across postnatal development. The presence of estrogen in the testicular fluid was indirectly ascertained by immunohistochemical detection of the P450 aromatase in the testis. There was no immunohistochemically detectable difference in the expression of P450 aromatase in the testes and ER${\beta}$ in the ED of WT and ${\alpha}$ERKO mice. ER${\alpha}$ was only detected in the ED of WT mice. The absence of ER${\alpha}$ in the ED of postnatally developing mice resulted in differential expression of mRNAs and/or proteins for carbonic anhydrase II, $Na^+/H^+$ exchanger 3, down-regulated in adenoma, cystic fibrosis transmembrane regulator, and $Na^+/K^+$ ATPase ${\alpha}$. Our data indicate that the absence of ER${\alpha}$ resulted in altered expression of an epithelial ion producer and transporters during postnatal development of mice. We conclude that the presence of ER${\alpha}$is important for regulation of the ED function during the prepubertal developmental and postpubertal period.

Functional analysis of SH3 domain containing ring finger 2 during the myogenic differentiation of quail myoblast cells

  • Kim, Si Won;Lee, Jeong Hyo;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1183-1189
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    • 2017
  • Objective: Owing to the public availability of complete genome sequences, including avian species, massive bioinformatics analyses may be conducted for computational gene prediction and the identification of gene regulatory networks through various informatics tools. However, to evaluate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite. Methods: Due to a lack of quail genomic sequence information, we first identified the partial genomic structure and sequences of the quail SH3 domain containing ring finger 2 (SH3RF2) gene. Subsequently, SH3RF2 was knocked out using clustered regularly interspaced short palindromic repeat/Cas9 technology and single cell-derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation. Results: Through a T7 endonuclease I assay and genotyping analysis, we established an SH3RF2 knockout (KO) QM7#4 subline with 61 and 155 nucleotide deletion mutations in SH3RF2. After the induction of myotube differentiation, the expression profiles were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis. Conclusion: We did not detect any statistically significant role of SH3RF2 during myotube differentiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.

Differential Expression of $PKD2$-Associated Genes in Autosomal Dominant Polycystic Kidney Disease

  • Yook, Yeon-Joo;Woo, Yu-Mi;Yang, Moon-Hee;Ko, Je-Yeong;Kim, Bo-Hye;Lee, Eun-Ji;Chang, Eun-Sun;Lee, Min-Joo;Lee, Sun-Young;Park, Jong-Hoon
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.16-22
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    • 2012
  • Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture. The proteins encoded by the $PKD1$ and $PKD2$ genes, mutations in which account for nearly all cases of ADPKD, may help guard against cystogenesis. Previously developed mouse models of $PKD1$ and $PKD2$ demonstrated an embryonic lethal phenotype and massive cyst formation in the kidney, indicating that $PKD1$ and $PKD2$ probably play important roles during normal renal tubular development. However, their precise role in development and the cellular mechanisms of cyst formation induced by $PKD1$ and $PKD2$ mutations are not fully understood. To address this question, we presently created $Pkd2$ knockout and $PKD2$ transgenic mouse embryo fibroblasts. We used a mouse oligonucleotide microarray to identify messenger RNAs whose expression was altered by the overexpression of the $PKD2$ or knockout of the $Pkd2$. The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction. Herein, we confirmed differential expressions of several genes including aquaporin-1, according to different $PKD2$ expression levels in ADPKD mouse models, through microarray analysis. These data may be helpful in $PKD2$-related mechanisms of ADPKD pathogenesis.