• Title/Summary/Keyword: noncoding DNA

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Cloning and sequencing of the gene encoding structural protein of canine parvovirus isolated in Korea (국내에서 분리된 canine parvovirus의 구조유전자 cloning과 염기서열 분석)

  • Park, Jong-hyeon;Song, Jae-young;Lee, Jung-bok;Hyun, Bang-hun;An, Soo-hwan;Jun, Moo-hyung
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.555-567
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    • 1992
  • In this study gene encoding structural proteins of a CPV isolate was cloned and sequenced to elucidate the molecular genetical properties of the canine parvoviruses isolated from the field. Six recombinant plasmids of pEP3, p1471, p2070, pEP069, pEP338 and p14711p were constructed from the map positions 22 to 98 of RF DNA to clone the VP1 and VP2 genes of CPV-V20. Sequentialy the gene comprising 3780 nucleotides were sequenced by dideoxy chain termination method. When nucleotide sequence of gene encoding the structural proteins of CPV-V20 was compared with those of other strains, CPV-N, CPV-d and CPV-780929 published previously, DNA, homologies to CPV-V20 were 99.87% with CPV-N, 99.73% with CPV-d, 96.85% with CPV-780929 and 98.4% with FPLV-Carl, respectively. The DNA sequence data of CPV-V20 showed seven point mutations and also deletion of 135 nucleotides from the nucleotide position 4745 to 4879 located in the 3'-noncoding region of CPV-N.

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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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Distribution of Length Variation of the mtDNA 9-bp Motif in the Intergenic COII/tRNAX$^{Lys}$ Region in East Asian Populations

  • Han Jun Jin;Jeon Won Choi;Dong Jik Shin;Jung Min Kim;Wook Kim
    • Animal cells and systems
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    • v.3 no.4
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    • pp.393-397
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    • 1999
  • Length variations in human mitochondrial DNA (mtDNA) offer useful markers in the study of female aspects of human population history. One such length variation is a 9-bp deletion in the small noncoding segment located between the COII and Iysine tRNA genes (COII/tRNA/$^{Lys}$ intergenic region) which usually contain two tandemly arranged copies of a 9-bp sequence (ccccctcta) in human mtDNA. The mtDNA 9-bp deletion and polymorphic variants of expanded 9-bp repeat motif in the intergenic COII/tRNA$^{Lys}$ region have been found at varying frequencies among different human ethnic groups. We have examined the length variation of the mtDNA COII/tRNA$^{Lys}$ intergenic region from a total of 813 individuals in east Asian populations. The occurrence of the 9-bp deletion was found to be relatively homogeneous in northeast Asian populations (Chinese, 14.2%; Japanese, 14.3%: Koreans, 15.5%), with the exception of Mongolians (5.1%). In contrast, Indonesians (25.0%) and Vietnamese (23.2%) of the southeast Asian populations appeared to have relatively high frequencies of the 9-bp deletion. We identified the existence of a new expanded 9-bp repeat motif which likely resulted from a slipped mispairing insertion of six more cytosines in the intergenic COII$^{Lys}$ region. It was present at low frequencies in the Korean (2/349) and Japanese populations (2/147). Based on the results of this study, the Korean population may reflect a close genetic affinity with the Japanese and Chinese populations than the others surveyed east Asian populations.

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Comparison of Hybridization Behavior between Double and Single Strand of Targets and the Application of Asymmetric PCR Targets in cDNA Microarray

  • Wei, Qing;Liu, Sanzhen;Huang, Jianfeng;Mao, Xueying;Chu, Xiaohui;Wang, Yu;Qiu, Minyan;Mao, Yumin;Xie, Yi;Li, Yao
    • BMB Reports
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    • v.37 no.4
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    • pp.439-444
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    • 2004
  • Double stranded targets on the cDNA microarray contain representatives of both the coding and noncoding strands, which will introduce hybridization competition with probes. Here, the effect of double and single strands of targets on the signal intensity and the ratios of Cy5/Cy3 within the same slide were compared. The results show that single stranded targets can increase the hybridization efficiency without changing the Cy5/Cy3 ratio. Based on these results, a new strategy was established by generating cDNA targets with asymmetric PCR, instead of conventional PCR, to increase the sensitivity of the cDNA microarray. Furthermore, the feasibility of this approach was validated. The results indicate that the cDNA microarray system based on asymmetric PCR is more sensitive, with no decrease in the reliability and reproducibility as compared with that based on conventional symmetric PCR.

Molecular Characterization and Infectious cDNA Clone of a Korean Isolate of Pepper mild mottle virus from Pepper

  • Yoon, Ju-Yeon;Hong, Jin-Sung;Kim, Min-Jea;Ha, Ju-Hee;Choi, Gug-Seon;Choi, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.361-368
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    • 2005
  • A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were $96.9\%\;to\;100.0\%\;and\;97.5\%\;to\;98.6\%$, respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5'-end primer anchoring T7 RNA promoter sequence and 3'-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

Functional Enhancers As Master Regulators of Tissue-Specific Gene Regulation and Cancer Development

  • Ko, Je Yeong;Oh, Sumin;Yoo, Kyung Hyun
    • Molecules and Cells
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    • v.40 no.3
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    • pp.169-177
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    • 2017
  • Tissue-specific transcription is critical for normal development, and abnormalities causing undesirable gene expression may lead to diseases such as cancer. Such highly organized transcription is controlled by enhancers with specific DNA sequences recognized by transcription factors. Enhancers are associated with chromatin modifications that are distinct epigenetic features in a tissue-specific manner. Recently, super-enhancers comprising enhancer clusters co-occupied by lineage-specific factors have been identified in diverse cell types such as adipocytes, hair follicle stem cells, and mammary epithelial cells. In addition, noncoding RNAs, named eRNAs, are synthesized at super-enhancer regions before their target genes are transcribed. Many functional studies revealed that super-enhancers and eRNAs are essential for the regulation of tissue-specific gene expression. In this review, we summarize recent findings concerning enhancer function in tissue-specific gene regulation and cancer development.

First report of Cycas necrotic stunt virus from cultivated Daphne plants

  • Lee, B.Y.;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.148.1-148
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    • 2003
  • Natural virus infection of cultivated Daphe odora plants showing chlorosis and stunting was observed and their causal agent was investigated. An isolate of isometic virus was purified from infected leaf tissues, and it could infect systemic severe mosaic on Chenopodium quinoa and C. amaranticolor. cDNA library was generated from partially purified viral RNAs and oligo dT primer-pSPORTl system, and recombinant clones were selected and their inserts were sequenced randomly. Nucleotide sequences of the virus were analyzed by BLAST, and it was closely related to members of subgroup B in the genus Nepovirus. The sequence analysis suggest that the virus was identified as an isolate of Cycas necrotic stunt virus (CNSV) because it was 89.7 % and 94.7 % identical to known CNSV for the CP and 3' noncoding region, respecitively. RT-PCR was performed to screen disease incidence of CNSV in Daphe plants, and five out of 10 plants (50 %) were infected by CNSV This is the first sequence information of CNSV from Daphe plants.

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General Primer-Mediated PCR Detection of Enteroviruses Causing Aseptic Meningitis (General Primer를 이용한 무균성뇌막염 원인 바이러스 분석)

  • Kim, M.B.;Kim, K.S.;Bae, Y.B.;Song, C.Y.;Yoon, J.D.;Lee, K.H.;Shin, H.K.
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.215-225
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    • 1996
  • Aseptic meningits, an acute inflammation of the meninges, is a common illness during childhood. Virus is the most important cause of aseptic meningitis. Especially enterovirus causes approximately above 85% of all cases of aseptic meningitis. In 1993, there was a big epidemic of aseptic meningitis by ECHO 9 and ECHO 30 viruses. And ECHO 3 virus was isolated as a causative agent of aseptic meningitis in 1994. This study was aimed to detect the causative agent of aseptic meningitis in 1995 and to analyze the 5'-noncoding region which was used to detect virus. Virus was isolated from 87 stools and cerebrospinal fluid specimens of the patients by cultured RD and HEp-2 cell. Neutralizing antibody tests using enterovirus serum pool were performed on the specimens with cytopathic effect. 3 of ECHO 7 viruses and 5 of Coxsackie B3 viruses were isolated from stool specimens and 1 of ECHO 7 and Coxsackie B3 mixed type was confirmed from cerebrospinal fluid specimens. RNA was isolated from the culture supernatants of infected cells and general primers were selected in highly conserved part of the 5'-noncoding region of the enteroviral genome for RT-PCR. PCR product from this virus showed a 152bp band on gel electrophoresis. Sequence of obtained DNA was compared with prototype sequences by accessing to the Genebank database. 5'-noncoding region of isolated Coxsackie B3 virus, which has point mutations in nucleotide sequence positions 493, 497, 502, 523, was closely related to that of polio virus type 1, Mahoney strain. In case of isolated ECHO 7 virus, nucleotide has been changed from cytosine to thymine at position 581 and from thymine to cytosine at position 583. We concluded the causative agents of the outbreak of aseptic meningitis during June to July in 1995 were both ECHO 7 and Coxsackie B3 virus, and the primer used in this study could allow a rapid diagnosis of enteroviruses by PCR.

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Taxonomic position and genetic differentiation of Korean Astragalus mongholicus Bunge (한국산 황기의 분류학적 위치 및 유전적 분화)

  • Choi, In-Su;Kim, So-Young;Choi, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.43 no.1
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    • pp.12-21
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    • 2013
  • To clarify the taxonomic position for Astragalus nakaianus and provide correct scientific name for A. mongholicus cultivar in South Korea, we examined external morphological characters and sequence variations from ITS and five cp non-coding DNA regions. Genetic structure was also analyzed for 61 individuals from three populations using nine microsatellite loci. We found no significant difference between the South Korean cultivar and A. mongholicus var. dahuricus when morphology and ITS sequences were considered. Morphologically, A. nakaianus specimens varied somewhat from A. mongholicus var. mongholicus and var. dahuricus in habit, plant height, and lengths of leaf axis and leaflet. Although sequence data from ITS and cp noncoding DNA regions could not distinguished A. nakaianus from A. mongholicus, microsatellite analysis revealed strong structuring between the cultivar and A. nakaianus. Therefore, we conclude that the South Korean A. mongholicus cultivar should be treated as A. mongholicus var. dahuricus and that A. nakaianus should be merged into A. mongholicus as a variety, i.e., A. mongholicus var. nakaianus.

Complete Chloroplast DNA Sequence from a Korean Endemic Genus, Megaleranthis saniculifolia, and Its Evolutionary Implications

  • Kim, Young-Kyu;Park, Chong-wook;Kim, Ki-Joong
    • Molecules and Cells
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    • v.27 no.3
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    • pp.365-381
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    • 2009
  • The chloroplast DNA sequences of Megaleranthis saniculifolia, an endemic and monotypic endangered plant species, were completed in this study (GenBank FJ597983). The genome is 159,924 bp in length. It harbors a pair of IR regions consisting of 26,608 bp each. The lengths of the LSC and SSC regions are 88,326 bp and 18,382 bp, respectively. The structural organizations, gene and intron contents, gene orders, AT contents, codon usages, and transcription units of the Megaleranthis chloroplast genome are similar to those of typical land plant cp DNAs. However, the detailed features of Megaleranthis chloroplast genomes are substantially different from that of Ranunculus, which belongs to the same family, the Ranunculaceae. First, the Megaleranthis cp DNA was 4,797 bp longer than that of Ranunculus due to an expanded IR region into the SSC region and duplicated sequence elements in several spacer regions of the Megaleranthis cp genome. Second, the chloroplast genomes of Megaleranthis and Ranunculus evidence 5.6% sequence divergence in the coding regions, 8.9% sequence divergence in the intron regions, and 18.7% sequence divergence in the intergenic spacer regions, respectively. In both the coding and noncoding regions, average nucleotide substitution rates differed markedly, depending on the genome position. Our data strongly implicate the positional effects of the evolutionary modes of chloroplast genes. The genes evidencing higher levels of base substitutions also have higher incidences of indel mutations and low Ka/Ks ratios. A total of 54 simple sequence repeat loci were identified from the Megaleranthis cp genome. The existence of rich cp SSR loci in the Megaleranthis cp genome provides a rare opportunity to study the population genetic structures of this endangered species. Our phylogenetic trees based on the two independent markers, the nuclear ITS and chloroplast MatK sequences, strongly support the inclusion of the Megaleranthis to the Trollius. Therefore, our molecular trees support Ohwi's original treatment of Megaleranthis saniculifolia to Trollius chosenensis Ohwi.