• Title/Summary/Keyword: X mRNA

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Karyotypic Analysis and Physical Mapping of rRNA Gene Loci in Persicaria tinctoria (쪽의 핵형분석과 rRNA 유전자의 염색체상 위치)

  • Choi, Hae-Woon;Lee, Sang-Hoon;Kim, Soo-Young;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.3
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    • pp.195-198
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    • 2008
  • Karyotypic analysis and FISH (fluorescence in situ hybridization) with 45S and 5S rRNA genes were carried out in Persicaria tinctoria H Gross. The somatic metaphase chromosomes were ranged from 2.25 ${\mu}m$ to 1.50 ${\mu}m$ in length. Chromosome number was 2n = 4x = 40 with the basic number of x = 10. The chromosome complement of the species consisted of 16 pairs of metacentrics (chromososomes 1,2,3,4,6,7,8,9, 10, 11, 12, 13, 15, 18, 19 and 20) and 4 pairs of submetacentrics (chromosome 5, 14, 16 and 17). The karyotype formula was K(2n) = 4x = 32 m + 8 sm. In FISH analysis, three pairs of 45S rRNA gene loci on the terminal region of submetacentrics (chromosomes 5, 16 and 17) and two pairs of 5S rRNA gene loci on the centromeric region of metacentrics (chromosomes 9 and 11) were detected, respectively.

Association Analysis of Myosin Heavy-chain Genes mRNA Transcription with the Corresponding Proteins Expression of Longissimus Muscle in Growing Pigs

  • Men, X.M.;Deng, B.;Tao, X.;Qi, K.K.;Xu, Zi Wei
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.457-463
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    • 2016
  • The goal of this work was to investigate the correlations between MyHC mRNA transcription and their corresponding protein expressions in porcine longissimus muscle (LM) during postnatal growth of pigs. Five DLY ($Duroc{\times}Landrace{\times}Yorkshire$) crossbred pigs were selected, slaughtered and sampled at postnatal 7, 30, 60, 120, and 180 days, respectively. Each muscle was subjected to quantity MyHCs protein contents through an indirect enzyme-linked immunosorbent assay (ELISA), to quantity myosin heavy-chains (MyHCs) mRNA abundances using real-time polymerase chain reaction. We calculated the proportion (%) of each MyHC to total of four MyHC for two levels, respectively. Moreover, the activities of several key energy metabolism enzymes were determined in LM. The result showed that mRNA transcription and protein expression of MyHC I, IIa, IIx and IIb in LM all presented some obvious changes with postnatal aging of pigs, especially at the early stage after birth, and their mRNA transcriptions were easy to be influenced than their protein expressions. The relative proportion of each MyHC mRNA was significantly positively related to that of its corresponding protein (p<0.01), and MyHC I mRNA proportion was positively correlated with creatine kinase (CK), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) activities (p<0.05). These data suggested that MyHC mRNA transcription can be used to reflect MyHC expression, metabolism property and adaptive plasticity of porcine skeletal muscles, and MyHC mRNA composition could be a molecular index reflecting muscle fiber type characteristics.

Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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3'-terminal sequence of mucor racemosus 18S rRNA gene (Mucor racemosus 18S rRNA gene의 3'말단 염기해독)

  • 지근억;김진경
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.284-289
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    • 1991
  • the nucleotide sequence of the 3' terminal 568 bases of the 18S rRNA gene from Mucor racemosus was determined. The 3' end of the structural gene was identified by comparison with the published sequence for the Saccharomyces cerevisiae gene. The M. racemosus gene was found to share 83.8% homology with that of S. cerevisiae and 71-81% homology with those of human, mouse, maize, Xenopus laevis and Tetrahymena thermophila. The known methylation sites in X. laevis and human were also highly conserved in M. racemosus and located within most conserved regions of 18S RNA gene throughout evolution.

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Construction of spSac3 Null Mutants Defective in mRNA Export (mRNA의 핵에서 세포질로의 이동에 관여하는 spSac3 유전자의 결실돌연변이 제조와 특성 조사)

  • Kang Sook-Hee;Yoon Jin-Ho
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.153-155
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    • 2006
  • We constructed the null mutants of fission yeast Schizosaccharomyces pombe spSac3 gene that is homologous to budding yeast Saccharomyces cerevisiae SAC3 involved in mRNA export out of nucleus. Tetrad analysis showed that the spSac3 is essential for vegetative growth. The spSac3 mutants harboring pREP81X-spSac3 plasmid showed poly(A)+ RNA export defect in the presence of thiamine. These results suggest that spSac3 is also involved in mRNA export from the nucleus.

Protein Kinase C-$\beta$ Is Induced In Ionizing Irradiation Induced Pigmentation

  • Nelly Rubeiz;Park, Dee-Young;Barbara A. Gilchrest
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.209-212
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    • 2002
  • Cutaneous hyperpigmentation is a well-known consequence of both acute and chronic X-irradiation, although the molecular mechanisms involved are not well understood. Recently, protein kinase C-$\beta$ (PKC-$\beta$) was shown to activate tyrosinase, a key and the rate-limiting enzyme in melanogenesis [1]. In this study, we have investigated its role in mediating ionizing radiation-induced pigmentation by exposing cultured human melanocytes to X-irradiation. Increased tyrosinase activity after the 4 Gys exposure was observed within 48 hrs and total melanin content doubled after 7 days. Interestingly, tyrosinase mRNA level was not affected by X-irradiation. However, there was a 2-3 fold increase in PKC-$\beta$ mRNA after 48 hours of irradiation, coinciding with the increase in tyrosinase activity. This induction was not due to non-specific heat generated during the irradiation because when melanocytes were incubated at 4$0^{\circ}C$, there was no induction of PKC-$\beta$ mRNA. Taken together, these data suggest that X-irradiation induces cutaneous hyperpigmentation, at least in part, by up-regulating the level of PKC-$\beta$.

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Some Tertiary Interactions in 5S rRNA from Xanthomonas celebensis (Xanthomonas celebensis 5S rRNA의 몇 가지 삼차상호작용)

  • Bongrae Cho;Yeonghoon Lee;Myung-Un Choi;Inwon Park
    • Journal of the Korean Chemical Society
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    • v.37 no.2
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    • pp.237-243
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    • 1993
  • The primary and secondary structure of the 5S rRNA isolated from Xanthomonas celebensis were determined by enzymatic and chemical degradation methods. It consists of 119 nucleotides and contains no modified nucleosides. As with the 5S rRNAs of X. maltophilia and X. citri, it contains an additional uridine residue on the 5'-terminus. Its secondary structure was almost identical to the models previously proposed by us for the 5S rRNA of two Xanthomonas species. Its secondary structure consists of five helices, five loops and two bulges. The tertiary interactions in the 5S rRNA molecule were analyzed by Fe(II)-EDTA treatment and hybridization method using deoxyhexamer. From the fact that some adenine residues in loop M, region $I_1-C$, loop $H_1$, and loop $H_2$ become susceptible to diethylpyrocarbonate when the 5S rRNA was hybridized with deoxyhexamer complementary to the sequence $U_{35}CCCAU_{40}$ and that some nucleotide residues in loop M, loop $H_1$ and region $D-I_2$ become resistant Fe(II)-EDTA cleavage in the presence of $Mg^{2+}$, it is presumed that loops $H_1$ and $H_2$ interact with loop M in some way. In the tertiary interaction, the regions $I_1-C$ and $D-I_2$ seem to act as hinges in folding the stems $B-I_1-C$ and $D-I_2-E.$ It was found that loop $H_1$ changes into a smaller loop of three bases by forming noncanonical A : C base-pairs ih acidic environment.

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Quantitative comparison of mRNA expression of glucosyltransferase (GTF) between $xylitol-resistant(X^R)$ and $xylitol-sensitive(X^S)$ mutans streptococci (Mutans streptococci의 자일리톨 내성균주와 감성균주의 glucosyltransferase mRNA의 정량적 비교 연구)

  • Lee, MI-Na;Kim, Young-Jae;Lee, Sung-Hoon;Kim, Chong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.1
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    • pp.77-84
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    • 2006
  • Since the long-term exposure of mutans streptococci to xylitol is known to select for xylitol-resistant $(X^R)$ natural mutants, the occurrence and survival of such $(X^R)$ strains were performed in batch culture methods. The aim of the study was to compare the differentiation and quantification of mRNA expression of the gtf genes of $X^R\;and\;X^S$ mutans streptococci. Using a real-time reverse-transcription polymerase chain reaction, the expression of each gtf was determined. In $X^R$ strains, the relative levels of transcription of gtfB and gtfC were decreased while that of gtfD was increased, suggesting the presence of independent promoters. It also suggested that mutation related to production of glucosyltransferase occurred under the exposure of xylitol could explain the caries-preventive mechanisms of xylitol.

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Inhibition of Hepatitis B Virus Replication by in vitro Synthesized RNA

  • Yang, Yeon-Ju;Heo, Young-Shin;Kim, Jeong-Ki;Kim, Sang-Yong;Ahn, Jeong-Keun
    • Bulletin of the Korean Chemical Society
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    • v.26 no.9
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    • pp.1385-1389
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    • 2005
  • Human hepatitis B virus (HBV) is a pathogen related to the development of liver diseases including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). However, the efficient methods to suppress HBV replication have not been developed yet. Therefore, we have used RNA interference (RNAi) as a potential tool for the suppression of HBV replication. Here, we designed a 21 nt small intefering dsRNA (siRNA) against hepatitis B virus X (HBx) RNA with 3' overhanging ends derived from T7 promoter. It has been reported that HBV X protein plays an important role in HBV gene expression and viral replication. The suppression of HBx gene expression by the 21 nt siRNA was investigated by Northern blot analysis and chloramphenicol acetyl transferase (CAT) assay. The level of HBx mRNA was decreased by siRNA in a dose-dependent manner. We also found that the 21 nt siRNA inhibited the HBV replication in hepatocellular carcinoma cell.

Cytokine mRNA Expression in the Small Intestine of Weanling Pigs Fed Diets Supplemented with Specialized Protein or Peptide Sources

  • Zhao, J.;Harper, A.F.;Webb, K.E. Jr.;Kuehn, L.A.;Gilbert, E.;Xiao, X.;Wong, E.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1800-1806
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    • 2008
  • Cytokines play a central role in the mucosal immune response and are involved in regulation of nutrient absorption, metabolism and animal growth. This study investigated the effect of diet manipulation with specialized protein or peptide sources on expression of cytokine (IL-1, IL-6, IL-10, and TNF-${\alpha}$) mRNA abundance in different intestinal regions and at different ages post-weaning in piglets. A total of 48 (17 days of age, $6.16{\pm}0.34kg\;BW$) weanling pigs were fed either a corn-soy/whey protein basal diet, the basal diet supplemented with spray-dried plasma protein (SDPP), or the basal diet supplemented with $Peptiva^{(R)}$, a hydrolyzed marine plant protein. A fourth treatment group was fed the SDPP diet, but the feed intake level was limited (SDPP-LF). Pigs were killed at 3 and 10 d, and intestinal cytokine mRNA was measured by real-time PCR using the relative quantification method. The SDPP-LF group exhibited an increased TNF-${\alpha}$ mRNA abundance compared with the ad libitum SDPP group (p<0.05). The TNF-${\alpha}$ and IL-10 mRNA abundance increased from the proximal to distal part of the intestine, and the mRNA abundance was greater (p<0.01) in the distal intestine as compared with the proximal and middle intestine. The cytokines IL-1-${\beta}$, IL-10 and TNF-${\alpha}$ mRNA abundance also increased from d3 to d10 postweaning (p<0.01). In summary, restricted feeding increased the TNF-${\alpha}$ mRNA abundance in the small intestine, however neither SDPP nor peptide supplementation affected cytokine mRNA expression. Abundance of mRNA for most cytokines examined in this study increased with age post-weaning, suggesting that during 10 d after weaning the mucosal immune system is still under development.