• Title/Summary/Keyword: DNA molecules

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Detection and Genomic Analysis of Viroid-like RNA Molecules Isolated from Korean Peonies (한국산 작약에서 분리한 바이로이드 유사 RNA 분자의 확인과 유전자 분석)

  • 정동수;김무인;이재열
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.113-117
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    • 1997
  • Low moleuclar weight (LMW) RNAs were isolated form Korean peonies which expressed symptoms of stunt and epinasty. The LMW plant RNAs were purified by Qiagen column chromatography which could separate viroid specific nucleic acid at differential salt concentration. After the inoculation of the purified RNAs from the peonies, the inoculated tomatoes (cv. Rutgers) expressed the symptoms of stunt and epinasty. Also the same molecular weight RNAs with viroid-like RNAs were isolated from the inoculated tomatoes. Double-stranded cDNA were synthesized by the methods of reverse transcription (RT) and polymerase chain reaction (PCR) with the purified RNA and primers. The same cDNAs associated with viroid-like RNAs wre cloned from the inoculated tomatoes. The cDNA has been sequenced and its 375-nucleotides were arranged into secondary structure. The cloned cDNA showed 47~54% homology compared with other viroids. The sequence homology of the cloned cDNA were partially high with plant genomic RNAs.

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Directed Alignment of DNA Molecule between the gold electrodes (금 전극위에 DNA 분자의 정렬에 관한 연구)

  • Hwang, Hyun Suk;Kim, Hyung Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.8
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    • pp.5586-5590
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    • 2015
  • In this paper, the directed alignment methode of the DNA molecule between the Au electrodes was suggested for the application of nano devices. To fabricate the nano device coated DNA, 2-Aminoethanthiol(AET) was coated on Au electrodes which was formed using photo-lithography process on $SiO_2/Si$ substrates. In general, the AET that was a positive charge with $NH^{3+}$ was strongly combined under the electrostatic interaction with DNA molecule which had to be a negative charge. The DNA molecules could be easily aligned between Au electrodes coated with AET. The structures of the DNA molecules were investigated using AFM(Atomic force microscope), they were changed from single types to bundle according to the AET concentrations.

Surface Polarity Dependent Solid-state Molecular Biological Manipulation with Immobilized DNA on a Gold Surface

  • Lee, Jiyoung;Kim, Jeong Hee
    • International Journal of Oral Biology
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    • v.37 no.4
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    • pp.181-188
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    • 2012
  • As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 ${\mu}M$. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.

ILLEGITIMATE RECOMBINATION MEDIATED BY MAMMALIAN DNA TOPOISOMERASE II

  • BAE YOUNG-SEUK;IKEDA HIDEO
    • Proceedings of the Microbiological Society of Korea Conference
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    • 1991.04a
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    • pp.82-96
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    • 1991
  • To understand the mechanism of illegitimate recombination in mammalian cells, we have examined the recombination role of DNA topoisomerase II (Topo II ). We found that purified calf thymus Topo II mediates recombination between two phage $\lambda$ DNA molecules in an in vitro system. The enzyme mainly produced a linear monomer recombinant DNA that can be packaged in vitro. Novobiocin and anti-calf thymus Topo II antibody inhibit this ATP-dependent recombination. The recombinant molecules contain duplications or deletion, and most crossovers take place between nonhomologous sequences of $\lambda$ DNA, as judged by the sequences of recombination junctions. In order to study the effects of Topo II on illegitimate recombination in mammalian cells, we have developed a new shuttle vector, pNKl, which contains three bacterial genes, amp(APR), galK and neo($Km^R$). Using this system, we have shown that a Topo II inhibitor, VM26, stimulated deletion formation in pNK1 DNA in monkey COS1 cells. Both in vitro and in vivo results suggest that Topo II participates in illegitimate recombination in mammalian cells.

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Effects of LED irradiation on the expression of apoptosis-related molecules in human SH-SY5Y neuroblastoma cells

  • Cho, Kyu-Seung;Ryu, Sun-Youl;Choi, Hong-Ran
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.1
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    • pp.1-10
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    • 2007
  • To verify the inhibitory or protective effects of light-emitting diode(LED) irradiation on apoptotic cell death induced by $CoCl_2$, human SH-SY5Y cells were treated with $CoCl_2$ and LED were used to irradiate the cells. In the cell viability assay, cells were died slowly from $50{\mu}M$ to $250{\mu}M$ and about 50% of cells died after 12 hours at $400{\mu}M$ of $CoCl_2$. The Diff-Quik staining revealed that cells showed condensation of DNA and blebbing of the cell membrane. The DNA fragmentation assay revealed the DNA fragmentation, which is another apoptosis marker, occurred in cells treated with $400{\mu}M$ $CoCl_2$ for 16 hours. In the western blot for HIF-$1{\alpha}$, HIF-$1{\alpha}$ was expressed after 3 hours from induction and peaked maximally at 16 hours. In the cell viability assay of the effects of LED irradiation (at 590 nm for 1 hour 20 minutes), the cells showed more proliferation (about 20%) than the control group. The RPA assay of various apoptosis-related molecules showed that pro-apoptosis molecules such as Bax, Bak, and Bid were upregulated in the $CoCl_2$ treatment group. This means that the apoptotic cell population was increased. However there was some significant changes in LED irradiated cells. In the $CoCl_2$-treated LED irradiation group, those molecules were down-regulated more than in the only $CoCl_2$-treated group. These results have shown that $CoCl_2$ may induce apoptotic cell death in human SH-SY5Y neuroblastoma cells. And LED irradiation has a positive effect on apoptotic cells by down-regulation of pro-apoptotic molecules.

Aptamers (nucleic acid ligands) for trypsin-like serine proteases

  • Gal, Sang-Wan;Jeong, Yong-Kee;Satoshi Nishikawa
    • Journal of Life Science
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    • v.12 no.1
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    • pp.14-18
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    • 2002
  • Subpopulations of nucleotides that bind specifically to a variety of proteins have been isolated from a population of random sequence RNA/DNA molecules. Roughly one in $10^{13}$ random sequence RNA/DNA molecules folds in such a way as to create a specific binding site for small ligands. Since the development of in vitro selection procedure, more than 50 nucleic acid ligands (aptamers) have been isolated. These molecules are very useful for the study of molecular recognition between nucleic acid and protein/organic compound. In addition to these basic studies this method gives us a dream to produce new drugs against several diseases. We focused on several aptamers which specifically binds to trypsin-like serine proteases (thrombin, human neutrophil elastase, activated protein C and NS3 protease of human hepatitis C virus) and want to introduce their structural characteristics and some functions.

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Version Space Learning with DNA Molecules (DNA 분자를 이용한 Version Space 학습)

  • 임희웅;장해만;채영규;유석인;장병탁
    • Proceedings of the Korean Information Science Society Conference
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    • 2002.10d
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    • pp.361-363
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    • 2002
  • Version space는 목표 개념이 속성 값에 대한 제한조건의 연언(conjunction)으로 표현될 수 있는 귀납적 개념학습에서 가설공간을 표현하기 위해 사용된다. Version space의 크기는 속성 값의 수에 대해 지수적으로 증가하는데, 우리는 DNA 분자를 이용하여 version space를 표현하는 효율적인 방법을 제시한다. 또한 version space를 유지하기 위한 기본 연산과, 이를 DNA 분자를 이용하는 구현 방법이 제시된다. 또한 DNA 분자로 표현된 version space를 활용하여 새로운 example에 대한 분류를 예측하는 방법을 제시한다.

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Opposite Roles of B7.1 and CD28 Costimulatory Molecules for Protective Immunity against HSV-2 Challenge in a gD DNA Vaccine Model

  • Weiner, David B.;Sin, Jeong-Im
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.68-77
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    • 2005
  • Background: Costimulation is a critical process in Ag-specific immune responses. Both B7.1 and CD28 molecules have been reported to stimulate T cell responses during antigen presentation. Therefore, we tested whether Ag-specific immune responses as well as protective immunity are influenced by coinjecting with B7.1 and CD28 cDNAs in a mouse HSV-2 challenge model system. Methods: ELISA was used to detect levels of antibodies, cytokines and chemokines while thymidine incorporation assay was used to evaluate T cell proliferation levels. Results: Ag-specific antibody responses were enhanced by CD28 coinjection but not by B7.1 coinjection. Furthermore, CD28 coinjection increased IgG1 production to a significant level, as compared to pgD+pcDNA3, suggesting that CD28 drives Th2 type responses. In contrast, B7.1 coinjection showed the opposite, suggesting a Th1 bias. B7.1 coinjection also enhanced Ag-specific Th cell proliferative responses as well as production of Th1 type cytokines and chemokines significantly higher than pgD+pcDNA3. However, CD28 coinjection decreased Ag-specific Th cell proliferative responses as well as production of Th1 types of cytokines and chemokine significantly lower than pgD+pcDNA3. Only MCP-1 production was enhanced by CD28. B7.1 coimmunized animals exhibited an enhanced survival rate as well as decreased herpetic lesion formation, as compared to pgD+pcDNA3. In contrast, CD28 vaccinated animals exhibited decreased survival from lethal challenge. Conclusion: This study shows that B7.1 enhances protective Th1 type cellular immunity against HSV-2 challenge while CD28 drives a more detrimental Th2 type immunity against HSV-2 challenge, supporting an opposite role of B7.1 and CD28 in Ag-specific immune responses to a Th1 vs Th2 type.

Long non-coding RNA: its evolutionary relics and biological implications in mammals: a review

  • Dhanoa, Jasdeep Kaur;Sethi, Ram Saran;Verma, Ramneek;Arora, Jaspreet Singh;Mukhopadhyay, Chandra Sekhar
    • Journal of Animal Science and Technology
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    • v.60 no.10
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    • pp.25.1-25.10
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    • 2018
  • The central dogma of gene expression propounds that DNA is transcribed to mRNA and finally gets translated into protein. Only 2-3% of the genomic DNA is transcribed to protein-coding mRNA. Interestingly, only a further minuscule part of genomic DNA encodes for long non-coding RNAs (lncRNAs) which are characteristically more than 200 nucleotides long and can be transcribed from both protein-coding (e.g. H19 and TUG1) as well as non-coding DNA by RNA polymerase II. The lncRNAs do not have open reading frames (with some exceptions), 3`-untranslated regions (3'-UTRs) and necessarily these RNAs lack any translation-termination regions, however, these can be spliced, capped and polyadenylated as mRNA molecules. The flexibility of lncRNAs confers them specific 3D-conformations that eventually enable the lncRNAs to interact with proteins, DNA or other RNA molecules via base pairing or by forming networks. The lncRNAs play a major role in gene regulation, cell differentiation, cancer cell invasion and metastasis and chromatin remodeling. Deregulation of lncRNA is also responsible for numerous diseases in mammals. Various studies have revealed their significance as biomarkers for prognosis and diagnosis of cancer. The aim of this review is to overview the salient features, evolution, biogenesis and biological importance of these molecules in the mammalian system.

Expressional Profiling of Molecules Associated with Epigenetic Methylation-Related Fertility in the Rat Testis during Postnatal Period

  • Seo, Hee-Jung;Lee, Seong-Kyu;Baik, Haing-Woon;Lee, Ki-Ho
    • Journal of Animal Science and Technology
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    • v.54 no.3
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    • pp.157-163
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    • 2012
  • The male reproduction is precisely controlled by a number of intrinsic and extrinsic factors. These factors usually involve in expressional regulation of various molecules influencing on sperm production in the testis. A number of ways are employed to control the transcription of specific genes, including epigenetic modifications of DNA and histone molecules. DNA methylation of CpG dinucleotides is a commonly used regulatory mechanism for testicular genes associated with the fertility. Previous studies have demonstrated the infertility induced by improper DNA methylation of these genes. In the present research, we attempted to determine transcriptional expression of some of these genes in the rat testis at different postnatal ages using real-time PCR analysis. These genes include neurotrophin 3 (Ntf3), insulin-like growth factor II (Igf2), JmjC-domain-containing histone demethylase 2A 1 (Jhm2da), paired box 8 transcription factor (Pax8), small nuclear ribonucleoprotein polypeptide N (Snrpn), and 5,10-methylenetetrahydrofolate reductase (Mthfr). The expression levels of Ntf3, Igf2, and Snrpn genes were the highest at the neonatal age, followed by transient decreases at the prepubertal age. Expression of Jhm2da and Mthfr genes were continuously increased from the neonate to 1 year of age. The levels of Pax8 mRNA at the early ages were higher than those at the later ages of postnatal development. These findings suggest that expression of some fertility-associated testicular genes in the rat during postnatal period could be differentially regulated by the control of the degree of DNA methylation.