• 제목/요약/키워드: 특이 mRNA

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Nucleotide Sequences of Bovine Ornithine Decarboxylase mRNA (젖소 Ornithine Decarboxylase mRNA의 염기서열)

  • Sung, Chang;Sparks, Robert
    • Korean Journal of Agricultural Science
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    • v.20 no.2
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    • pp.189-200
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    • 1993
  • Ornithine decarboxylase is the first and rate limiting enzyme in the biosynthesis of polyamines in mammalian cells. During cell growth the enzyme is regulated by rapid changes in the level of its mRNA and protein. To explore the molecular basis of these changes, ODC-specific complementary DNA (cDNA) clones were isolated from a bovine cDNA library. This region of the cDNA contained a portion of the open reading frame, a 3'noncoding region, and a poly-A tail of 456, 348, and 14 nucleotides, respectively. A comparison of the deduced sequence of the carboxyl terminal 151 amino acids of ODC with amino acid sequences in the same region of the enzyme from human, mouse, rat, and hamster showed greater than 88% identity in these proteins. The highly conserved nature of the amino acid sequences may be related to the important role of ODC in cell growth and differentiation.

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Development and Characterization of RNA Aptamers for Phosphorylated Amino Acids (인산화 형태의 아미노산들에 대한 엡타머의 개발과 특성연구)

  • Cho Suhyung;Kim Byung-Gee
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.88-92
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    • 2005
  • Phosphorylation of amino acid residues in proteins, plays a major role in biological mechanism. Phosphorylation acts as a process regulating the protein activity in variable pathways such as metabolism, signal transduction and cell division. Therefore the development of ligands for phosphoamino acids are an important work for protein analysis and proteomics studies. In this study, RNA aptamers for o-phosphoserine, o-phosphotyrosine and o-phosphotyrosine which appears frequently in nature were developed by in vitro evolution method. We could obtain similar sequences from random RNAs of 40 mer by SELEX method through 10 cycles. As result, the aptamers for o-phosphoserine and o-phosphothreonine among phosphoamino acids aptamers showed high affinity of Kd=2.60 nM and 2.65 nM for their target molecules, respectively. In addition, these aptamers could be confirmed the high selectivity for their target.

Increased mRNA Related Ovarian Maturation during Induction of Maturational Competence in Red Seabream, Pagrus major (참돔, Pagrus major의 성숙능력 유도시 증가된 난성숙 관련 mRNA)

  • Choi, Cheol-Young;Chang, Young-Jin;Takashima, Fumio
    • Development and Reproduction
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    • v.4 no.1
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    • pp.125-131
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    • 2000
  • This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

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Induced Expression of Doublesex-and mab-3-related Transcription Factor-1 (DMRT-1) mRNA by Testosterone in the Olive Flounder, Paralichthys olivaceus ovary (Testosterone 처리에 의한 넙치, Paralichthys olivaceus 난소에서 doublesex-and mab-3-related transcription factor-1 (DMRT-1) mRNA의 발현 유도)

  • Jo, Pil-Gue;An, Kwang-Wook;Kim, Na-Na;Choi, Yong-Ki;Cho, Sung-Hwoan;Min, Byung-Hwa;Lim, Han-Kyu;Choi, Cheol-Young
    • Journal of Aquaculture
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    • v.20 no.3
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    • pp.199-202
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    • 2007
  • We isolated a 317 bp of partial cDNA for doublesex-and mab-3-related transcription factor-1 (DMRT-1) from the testis of olive flounder, Paralichthys olivaceus using RT-PCR. Based on the multiple sequence alignment, olive flounder DMRT-1 shared relatively high sequence homology (82 to 94%) with orthologues from other teleost species such as Atlantic halibut, Hippoglossus hippoglossus, black porgy, Acanthopagrus schlegeli and rainbow trout, Oncorhynchus mykiss. DMRT-1 mRNA was predominantly expressed in the testis of olive flounder. In our investigation for the effect of testosterone treatment in vivo on induced expression of ovarian DMRT-1 transcript, mRNA levels of DMRT-1 in ovary were significantly up-regulated by testosterone treatments (0.3 or $3.0{\mu}g$ testosterone/g body weight for 12 to 36 hours) as judged by RT-PCR analysis. In overall, transcriptional stimulation of DMRT-1 during treatments was more affected by doses of testosterone than treatment durations. This result strongly suggests that the regulation of DMRT-1 be tissue- and gender-specific in olive flounder, and also provides useful baseline knowledge on the testosterone-mediated regulation in the reproductive physiology of this species.

Genomic Variation and Toxin Specificity of Ustilago maydis Viruses from Progeny Strains as a Result of Artificial Mating (Ustilago maydis의 Mating 과정에 따른 Virus 유전자의 변이에 관한 연구)

  • 강인식;이세원
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.105-110
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    • 1997
  • Ustilago maydis strains (A-series and SH-series) containg virus or viral dsRNAs were artificially mated in corn seedling to generate 6 progeny strains, designated A23, A45, A21l, A31O, SH24 and SH61O. The dsRNA patterns of progeny strains were identical to those of the parental strains and there was no molecular exclusion mechanism among dsRNAs of parental strains. Virus particles were purified from 6 progeny strains and viral dsRNAs were analyzed on 5% PAGE. There was no mixed encapsidation between virus or dsRNAs of parental strains. Progeny strain SH6l4 produced toxin which inhibits the growth of SH9, SHIO and SH11. Likewise, toxins from A310 and SH24 inhibited growth of the SH11 strains. These results indicate that the presence of different types of dsRNA does not interfere the expression of toxin gene.

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Expression profile of defense-related genes in response to gamma radiation stress (방사선 스트레스 반응 방어 유전자의 탐색 및 발현 분석)

  • Park, Nuri;Ha, Hye-Jeong;Subburaj, Saminathan;Choi, Seo-Hee;Jeon, Yongsam;Jin, Yong-Tae;Tu, Luhua;Kumari, Shipra;Lee, Geung-Joo
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.359-366
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    • 2016
  • Tradescantia is a perennial plant in the family of Commelinaceae. It is known to be sensitive to radiation. In this study, Tradescantia BNL 4430 was irradiated with gamma radiation at doses of 50 to 1,000 mGy in a phytotron equipped with a $^{60}Co$ radiation source at Korea Atomic Energy Research Institute, Korea. At 13 days after irradiation, we extracted RNA from irradiated floral tissues for RNA-seq. Transcriptome assembly produced a total of 77, 326 unique transcripts. In plantlets exposed to 50, 250, 500, and 1000 mGy, the numbers of up-regulated genes with more than 2-fold of expression compared that in the control were 116, 222, 246, and 308, respectively. Most of the up-regulated genes induced by 50 mGy were heat shock proteins (HSPs) such as HSP 70, indicating that protein misfolding, aggregation, and translocation might have occurred during radiation stress. Similarly, highly up-regulated transcripts of the IQ-domain 6 were induced by 250 mGy, KAR-UP oxidoreductase 1 was induced by 500 mGy, and zinc transporter 1 precursor was induced by 1000 mGy. Reverse transcriptase (RT) PCR and quantitative real time PCR (qRT-PCR) further validated the increased mRNA expression levels of selected genes, consistent with DEG analysis results. However, 2.3 to 97- fold higher expression activities were induced by different doses of radiation based on qRT-PCR results. Results on the transcriptome of Tradescantia in response to radiation might provide unique identifiers to develop in situ monitoring kit for measuring radiation exposure around radiation facilities.

A Specific Biomarker for The Diagnosis of Equine Laminitis (말의 제엽염 진단을 위한 특이적 생물지표)

  • Lee, Seungwoo;Lee, Seung Heon;Kim, Myung-Chul;Kim, Yong-Baek;Ryu, Doug-Young
    • Journal of Veterinary Clinics
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    • v.32 no.1
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    • pp.62-68
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    • 2015
  • There is no reliable indicator available for the diagnosis of horse laminitis, although the disease is common and costly. This study was performed to develop a specific diagnostic biomarker for laminitis. We have identified 33 differentially expressed proteins in plasma of a horse suffering laminitis that is experimentally induced by an overdose of oligofructose, in comparison with normal horse plasma. Among the proteins, myosin-9 mRNA was found in RNA sequencing analysis to be expressed specifically in laminitis tissues compared to other horse tissues. It is thus suggested that expression of plasma myosin-9 may be used for the diagnosis of equine laminitis.

DNA Sequence and Characteristics of Muscle Development cDNA Clone Derived from Korean Native Chicken (재래닭의 근육 성장과 관련되는 cDNA Clone의 염기서열 및 특성)

  • Sun, S.S.;Myung, K.H.;Kuk, K.;Kim, N.O.
    • Korean Journal of Poultry Science
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    • v.33 no.4
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    • pp.249-254
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    • 2006
  • This experiment was conducted to examine the effective DNA related with muscle growth of Korean native chicken. cDNA library was constructed with mRNA subtraction from Korean native chicken to Cornish. Total mRNA was purified from pectoralis muscle of adult chicken. Five clones were compared their DNA sequence and characteristics based on GenBank. Clone NDS-1 (618nt) was low homology (10%) with other species, but it is closely related with triosephosphate isomerase which is play an important role in glycolysis. Clone NDS-6 (651nt) is corresponding to glyceraldehyde-3-phosphate dehydrogenase. These two clones are encoding to enzymes in key role in glycolysis. However, other three clones (NDS-2, NDS-10, NDS-12) have low homology with other species about 5.0%. These clones were not similar with any other eukaryotics. Therefore, three clones (NDS-2, NDS-10, NDS-12) are high possibility of specific DNA for muscle growth in Korean native chicken.

Transcriptional Response of Major Antioxidant Enzyme Genes to Heat Stress in Mud Loach (Misgurnus mizolepis) (고온 스트레스에 대한 미꾸라지(Misgurnus mizolepis) 항산화 효소 유전자들의 발현 특징)

  • Cho Young-Sun;Lee Sang-Yoon;Bang In-Chul;Kim Dong-Soo;Nam Yoon-Kwon
    • Journal of Aquaculture
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    • v.19 no.3
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    • pp.157-165
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    • 2006
  • Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.

Gene Expression of Surfactant Protein B and C in Endotoxin and Thiourea Treated Rats (내독소 및 Thiourea 투여 후 Surfactant protein B와 C 유전자 발현의 비교 관찰)

  • Sohn, Dong Hyun;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.510-521
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    • 2003
  • Background : The surfactant specific proteins, SP-B and SP-C are believed to be important regulators of the surfactant function and homeostasis. Since acute respiratory distress syndrome(ARDS) is usually viewed as the functional and morphological expression of a similar underlying lung injury caused by a variety of insults, and since abnormalities in the surfactant function have been described in ARDS, the authors investigated the different effects of endotoxin and thiourea on the accumulation of mRNA encoding SP-B and SP-C. Methods : Sprague-Dawley rats were given 5 mg/kg of an intraperitoneal endotoxin from Salmonella enteritidis and 3.5 mg/kg intraperitoneal thiourea and were sacrificed at different time periods. Results : 1. The SP-B mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 26.1% and 50%, respectively(P<0.01, P<0.001). 2. The SP-B mRNA levels 24 hours after the 3.5 mg/kg thiourea treatment was reduced by 9.8% and 12.5%, respectively. 3. The SP-C mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 38.7% and 53.6%, respectively(P<0.01, P<0.001). 4. The SP-C mRNA level 6 hours after the 3.5 mg/kg thiourea treatment was reduced by 22.8%(P<0.05). Conclusion : These results indicate that the differential regulation of the hydrophobic surfactant proteins in vivo is evident, and suggest that the hydrophobic surfactant proteins might be differentially regulated during lung injury at different time periods without altering the lung wet to dry ratios. The mechanism of these alternations at the different time periods and the different kinds of etiology remain to be determined.