• Title/Summary/Keyword: RNA profile

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Identification of Lactic Acid Bacteria from Meat by Low Molecular Weight(LMW) RNA Profiles (Low Molecular Weight(LMW) RNA Profiles에 의한 젖산균의 동정)

  • Cha, Woen-Suep
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.6
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    • pp.681-685
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    • 1992
  • Low molecular weight RNA(LMW RNA : 5S rRNA and tRNAs, <150 nucleotides) profiles of several bacteriocin production lactic acid bacteria from pig meats and reference lactic acid bacteria were generated on 10% denaturing polyacrylamide gel electrophoresis. Data evaluation including three molecular weight markers enabled the calculation of relative nucleotide units(RNU) for every band. Gels profiles and RNU evaluations were effective for identification of lactic acid bacteria species. LMW RNA profiles of lactic acid bacteria showed no variation in dependence on APT(All Purpose Tryptone Broth), TSB(Tryptic Soy Broth), MRS(Lactobacilli MRS Broth) different cultural medium.

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Purification of Radiolabeled RNA Using Sephadex G-15 or G-50 Chromatography (Sephadex G-15 또는 G-50 chromatography를 이용한 방사성 동위원소로 표지된 RNA의 정제)

  • Yoo, Beong-Gyu;Lee, Jong-Seok
    • Journal of radiological science and technology
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    • v.21 no.1
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    • pp.65-68
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    • 1998
  • We attempted to purify radiolabeled RNA using Sephadex G-15 and G-50 chromatography instead of commercial RNA purification kit. In the Sephadex G-15 chromatography the major portion of RNA was eluted in the fractions ranging from 3rd to 5th whereas broad elution profile of RNA was obtained from the Sephadex G-50 chromatography. The elution profile and purity of RNA obtained from Sephadex G-15 chromatography was very similar to that by commercial RNA purification kit. Furthermore, operating time required for purification of RNA by Sephadex G-15 was rather smaller than that by commercial kit. Overall results suggest that the purification of radiolabeled RNA using Sephadex G-15 is more money and time saying than using commercial RNA purification kit.

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The Increment of Purine Specific Sodium Nucleoside Cotransporter mRNA in Experimental Fibrotic Liver Induced by Bile Duct Ligation and Scission

  • Lee, Sung-Hee;Chae, Keon-Sang;Nan, Ji-Xing;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.613-619
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    • 2000
  • We investigated the expression profiles of rat fibrotic liver induced by bile duct ligation and scission (BDL/S) using the 3'-directed cDNA libraries. The possibility that the 3'-directed cDNA library represents the mRNA population faithfully was examined by northern blots. During the northern analysis based on fibrotic liver expression profile, we found for the first time that purine specific sodium nucleoside cotransporter (SPNT) was upregulated in BDL/S-induced fibrotic liver. To determine whether the accumulation of bile juice could affect the expression of SPNT mRNA or not, we examined the change of SPNT mRNA expression at 3, 14, 28 days after BDL/S operation. No change in SPNT expression was observed in rat liver at 3 days after surgery. In contrast, there were significant increases in SPNT expression at 14 and 28 days after surgery. We also examined whether chronic liver damage affected SPNT mRNA expression. SPNT mRNA level was significantly increased in BDL/S-induced fibrotic rat liver, whereas no significant change was obserbed in fibrotic livers chronically exposed to carbon tetrachloride or dimethylnitrosamine. From the above results, although further study might be needed, it was considered that the increment of SPNT mRNA in BDL/S liver morphological compatibility to human was remarkable.

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Analysis of Microbial Diversity in Nuruk Using PCR-DGGE (PCR-DGGE를 이용한 누룩에서의 미생물 다양성 분석)

  • Kwon, Seung-Jik;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.1
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    • pp.110-116
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    • 2012
  • Nuruk plays a significant role in the flavor and quality of Takju and Yakju, which are produced through saccharification and alcohol fermentation by various microorganisms. In this study, we identified microbial strains isolated from a plate count and PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes, in order to characterize bacterial and fungal diversity in Sansung Nuruk. The numbers of bacteria and fungi in Nuruk were $1.5{\times}10^9$ CFU/g and $2.2{\tims}10^8$ CFU/g, respectively. The 16S rRNA gene sequence indicated that the predominant bacteria in the isolates and PCR-DGGE profile of Nuruk were Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcus spp., Weissella spp., Staphylococcus spp., endophytic bacterium, uncultured Gamma-proteobacteria, uncultured Cyanobacteria, and Actinobacteria. Dominant bacteria from the PCR-DGGE profile were Pediococcous pentosaceus and uncultured Cyanobacteria. The 28S rRNA gene sequence indicated the predominant fungi in the isolates and PCR-DGGE profile to be Trichomonascus spp. Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp. Aspergillus spp., and Cladosporium spp. Dominant fungi from the PCR-DGGE profile were Pichia kudriavzevii and Aspergillus oryzae. The PCR-DGGE technique was used for the first time in this study to assess a microbial community in Nuruk and proved to be an effective protocol for profiling microbial diversity.

Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE (PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석)

  • Kwon, Seung-Jik;Ahn, Tae-Young;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.2
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    • pp.232-238
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    • 2012
  • Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

Charactrization of encephalomyocarditis virus isolated from Korea (국내에서 분리한 encephalomyocarditis virus의 성상)

  • Kim, Won-yong;Mah, Jum-sool;Kim, Chul-joong;Kang, Shien-young;Choi, Jae-yoon;Ha, Yong-kong
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.585-595
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    • 1992
  • We have investigated the characteristics of encephalomyocarditis(EMC) virus isolated in Korea. The CPE, buoyant density, polypeptide profile and the size of RNA of EMC virus were examined. The granulation, pyknosis and necrosis were observed from 30 to 48 hour's post inoculation of the virus into baby hamster kidney and lung cells. The buoyant density was 1.30 and $1.35g/m{\ell}$. Three different polypeptides, 26Kd, 32Kd, and 34Kd in size, were observed and the size of viral RNA was 7.7Kb.

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An RNA Mapping Strategy to Identify Ribozyme-Accessible Sites on the Catalytic Subunit of Mouse Telomerase

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • v.5 no.1
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    • pp.32-35
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    • 2007
  • Telomerase reverse transcriptase (TERT) is an enzymatic ribonucleoprotein that prolongs the replicative life span of cells by maintaining protective structures at the ends of eukaryotic chromosomes. Telomerase activity is highly up-regulated in 85-90% of human cancers, and is predominately regulated by hTERT expression. In contrast, most normal somatic tissues in humans express low or undetectable levels of telomerase activity. This expression profile identifies TERT as a potential anticancer target. By using an RNA mapping strategy based on a trans-splicing ribozyme library, we identified the regions of mouse TERT (mTERT) RNA that were accessible to ribozymes. We found that particularly accessible sites were present downstream of the AUG start codon. This mTERTspecific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors.

MicroRNA Expression Profile Analysis Reveals Diagnostic Biomarker for Human Prostate Cancer

  • Liu, Dong-Fu;Wu, Ji-Tao;Wang, Jian-Ming;Liu, Qing-Zuo;Gao, Zhen-Li;Liu, Yun-Xiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3313-3317
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    • 2012
  • Prostate cancer is a highly prevalent disease in older men of the western world. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression via posttranscriptional inhibition of protein synthesis. To identify the diagnostic potential of miRNAs in prostate cancer, we downloaded the miRNA expression profile of prostate cancer from the GEO database and analysed the differentially expressed miRNAs (DE-miRNAs) in prostate cancerous tissue compared to non-cancerous tissue. Then, the targets of these DE-miRNAs were extracted from the database and mapped to the STRING and KEGG databases for network construction and pathway enrichment analysis. We identified a total of 16 miRNAs that showed a significant differential expression in cancer samples. A total of 9 target genes corresponding to 3 DE-miRNAs were obtained. After network and pathway enrichment analysis, we finally demonstrated that miR-20 appears to play an important role in the regulation of prostate cancer onset. MiR-20 as single biomarker or in combination could be useful in the diagnosis of prostate cancer. We anticipate our study could provide the groundwork for further experiments.

How are Bayesian and Non-Parametric Methods Doing a Great Job in RNA-Seq Differential Expression Analysis? : A Review

  • Oh, Sunghee
    • Communications for Statistical Applications and Methods
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    • v.22 no.2
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    • pp.181-199
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    • 2015
  • In a short history, RNA-seq data have established a revolutionary tool to directly decode various scenarios occurring on whole genome-wide expression profiles in regards with differential expression at gene, transcript, isoform, and exon specific quantification, genetic and genomic mutations, and etc. RNA-seq technique has been rapidly replacing arrays with seq-based platform experimental settings by revealing a couple of advantages such as identification of alternative splicing and allelic specific expression. The remarkable characteristics of high-throughput large-scale expression profile in RNA-seq are lied on expression levels of read counts, structure of correlated samples and genes, larger number of genes compared to sample size, different sampling rates, inevitable systematic RNA-seq biases, and etc. In this study, we will comprehensively review how robust Bayesian and non-parametric methods have a better performance than classical statistical approaches by explicitly incorporating such intrinsic RNA-seq specific features with flexible and more appropriate assumptions and distributions in practice.

MicroRNA Analysis during Cultured Odontoblast Differentiation

  • Park, Min-Gyeong;Lee, Myoung-Hwa;Yu, Sun-Kyoung;Park, Eu-Teum;Kim, Seog;Lee, Seul-Ah;Moon, Yeon-Hee;Kim, Heung-Joong;Kim, Chun-Sung;Kim, Do-Kyung
    • International Journal of Oral Biology
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    • v.37 no.3
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    • pp.146-152
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    • 2012
  • MicroRNAs (miRNAs, miRs) are about 21-25 nucleotides in length and regulate mRNA translation by base pairing to partially complementary sites, predominantly in the 3'-untranslated region (3'-UTR) of the target mRNA. In this study, the expression profile of miRNAs was compared and analyzed for the establishment of miRNA-related odontoblast differentiation using MDPC-23 cells derived from mouse dental papilla cells. To determine the expression profile of miRNAs during the differentiation of MDPC-23 cells, we employed miRNA microarray analysis, quantitative real-time PCR (qRT-PCR) and Alizaline red-S staining. In the miRNA microarray analysis, 11 miRNAs were found to be up- or down-regulated more than 3-fold between day 0 (control) and day 5 of MDPC-23 cell differentiation among the 1,769 miRNAs examined. In qRT-PCR analysis, the expression levels of two of these molecules, miR-194 and miR-126, were increased and decreased in the control MDPC-23 cells compared with the MDPC-23 cells at day 5 of differentiation, respectively. Importantly, the overexpression of miR-194 significantly accelerated mineralization compared with the control cultures during the differentiation of MDPC-23 cells. These results suggest that the miR-194 augments MDPC-23 cell differentiation, and potently accelerates the mineralization process. Moreover, these in vitro results show that different miRNAs are deregulated during the differentiation of MDPC-23 cells, suggesting the involvement of these genes in the differentiation and mineralization of odontoblasts.