• Title/Summary/Keyword: oligodeoxyribonucleotide

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Synthesis of Dimeric Oligodeoxyribonucleotide (이합체성 올리고디옥시리보핵산의 합성)

  • Song, Jeong-Sup
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.8 no.6
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    • pp.1566-1571
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    • 2007
  • Oligodeoxyribonucleotide bearing the di-acetylenic linkage has been prepared. Staring from 5-Iodo-2'-deoxyuridine(1), a four-step sequence, consisting of the Pd(0)-catalyzed Heck-type C-C coupling with acetylenic group, protection of 5'-hydroxy group, generation of acetylenic hydrogen, and Glaser oxidative coupling reaction leads to the dimeric oligodeoxyribonucleotide(5).

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Development of New Protecting Groups for Guanine Residue in Oligodeoxyribonucleotide Synthesis

  • Byung Jo Moon;Kyung Lan Huh
    • Bulletin of the Korean Chemical Society
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    • v.12 no.2
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    • pp.196-199
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    • 1991
  • Attempts were made to develop new protecting groups for 1,6-lactam function of 2-N-acyl guanine in oligodeoxyribonucleotide synthesis. Several acyl groups, aryl groups, and carbamoyl groups were tested. Dimethylcarbamoyl and phenylacetyl groups are shown to be a good combination for guanine residue. 6-O-Di-methylcarbamoyl-2-N-pheylacetyl-2'-deoxy guanosine have been successfully used in the synthesis of d[AAGCTT], which is Hind Ⅲ recognition sequence.

Chemical Synthesis of Oligodeoxyribonucleotides Using Paper Disk as a Support (Paper Disk를 지지체로 이용한 Oligodeoxyribonucleotides 화학적 합성)

  • Bong-Oh RO;Sung-Jun Kim;Dae-Hyun Shin
    • Journal of the Korean Chemical Society
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    • v.37 no.3
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    • pp.355-363
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    • 1993
  • The two deoxyribonucleotides, 5'-d(GAATTCCGGCCA) and 5'-d(CGAGCTGTC), were synthesized on disks of Whatmann chromatography paper (3MM) by the phosphite-triester method. The average yield of the dodecamer and the nonamer that was measured by trityl analysis is 76.1${\%}$ and 86.5${\%}$ respectively. Separation of the synthesized oligodeoxyribonucleotides were purified by HPLC. The two oligodeoxymers'bases were quantified by HPLC with snake venom phosphodiesterase and bacterial alkaline phosphatase. The numbers of bases were equal to the designed ones. The advantage of using a paper disk as a support allows us to synthesize many different oligodeoxyribenucleotides during one synthesis. This method is the most convenient synthetic technique to synthesize oligodeoxyribonucleotides easily and inexpensively in a relatively short time.

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Chemical Synthesis of Oligodeoxyribonucleotide ; Improvement of Deoxyribonucleoside Phosphorylation and Dideoxyribonucleotide Synthesis (Oligodeoxyribonucleotide의 화학적 합성 ; Deoxyribonucleoside의 인산화와 이량체 합성 방법의 개선)

  • Sang Jik Lee;Byung Soo Song;Jong Dae Kim
    • Journal of the Korean Chemical Society
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    • v.31 no.1
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    • pp.84-93
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    • 1987
  • The study was done with a focus on making the optimum condition on phosphorylation of deoxyribonucleoside with o-chlorophenylphosphoroditriazole as a phosphorylating agent. The result showed that the addition of 5 volume % pyridine to the dioxane solution accelerated the rate of reaction to a great extent and turned out to nearly quantitative yields on phosphorylation. On the basis of this improvement of optimum reaction conditions, a more efficient method to synthesize all-protected dideoxyribonucleotide from N, 5-O-blocked deoxyribonucleoside was developed. The dodecamer with a Hind Ⅲ recognition site was readily synthesized from five different dimers which were prepared through the newly improved method.

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Chemical Synthesis of an Oligodeoxyribonucleotide (Oligodeoxyribonucleotide의 화학적 합성)

  • Byong-Soo Song;Lee Sang-Jik;Jong-Dae Kim
    • Journal of the Korean Chemical Society
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    • v.30 no.1
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    • pp.90-100
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    • 1986
  • Along with synthesizing a model oligodeoxyribonuclotide GpApApTpTpCp which had the EcoRI recognition site according to "Phosphotriester Method", reported methods for protecting functions, condensation, deprotection, purifying products, quantification and identification were evaluated and modified. It was realized that oligomers were efficiently synthesized by elongating by dimer unit per step of the condensation and the condensation yields were decreased with increased numbers of the residues. The problems of quantification/identification involved in the oligonucleotide synthesis using small amount of the reactants were solved by employing UV/IR spectrophotometry and hplc/tlc. It was also proved that a nucleotide having the exposed 5'-OH function was well synthesized by condensing a 5'-OH nucleotide and a phosphodiester nucleotide whose molar ratio was intendedly made to be 1 : 1.2 and then detritylating followed by washing with an aqueous solution of sodium bicarbonate. Spectrophotometric and chromatographic data of the nucleotides and their derivatives concerned in this synthetic work were prepared.

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Construction of an Expression Vector System with the GAP Promoter in Saccharomyces cerevisiae (효모, Saccharomyces cervisiae의 GAP 유전자를 이용한 발현 벡터계의 개발)

  • 황요일;서애란;심상국;정동효
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.568-574
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    • 1991
  • The cloned glyceraldehyde-3-phosphate dehydrogenase (GAP) gene of Saccharomyces cereviszae (Holland et al., 1983) has been characterized. Based on the communication, we have also cloned 2.1 kb CAP DNA fragment and modified this fragment as a portable promoter. Two yeast expression vectors, one is YCp type vector being maintained at low copy number (1 or 2) and the other is YEp type vector at high copy number, have been constructed with the GAP promoter and the PH05' gene as a reporter. Our plasrnids were introduc,ed into S. cerevisiae HY-1, which has been improved. The $Trp^+$ transformants expressed APase activity efficiently and showed high level of PH05' transcripts.

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Synthesis and Characterization of Oligonucleotides Containing Site-Specific Bulky $N^2$-Aralkylated Guanines and $N^6$-Aralkylated Adenines

  • Moon, Ki-Young;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.139-146
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    • 2000
  • 7- Bromomethylbenz[a]anthracene is a known mutagen and carcinogen. The two major DNA adducts produced by this carcinogen, i.e., $N^2$-(benz[a]anthracen-7-yl methyl)-2'-deoxyguanosine (2, b[a]$a^2$G) and $N^6$-(benz[a]anthracen-7-ylmethyl)-2'-deoxyadenosine (4, b[a]$a^6$/A), as wel 1 as the simpler benzylated analogs,$N^2$-benzyl-2'deoxyguanosine (1, $bn^2$G) and $N^6$-benzyl-2'-deoxyadenosine (3, $bn^6$/A), were prepared by direct aralkylation of 2'-deoxyguanosine and 2'-deoxyadenosine. To determine the site-specific mutagenicity of these bulky exocyclic amino-substituted adducts, the suitably protected nucleosides were incorporated into 16-base oligodeoxyribonucleotides in place of a normal guanine or adenine residues which respectively are part of the ATG initiation codon for the lac Z' \alpha-complementation gene by using an in situ activation approach and automated phosphite triester synthetic methods. The base composition and the incorporation of the bulky adducts into synthetic oligonucleotides were characterized after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis.

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Relationships of the Lithium-Induced Growth Inhibition of C6 Rat Glioma Cell to Expression of the Insulin-like Growth Factor System Components (C6 Rat Glioma Cell에서 리튬에 의한 성장 억제와 Insulin-like Growth Factor System Components의 발현과의 관계)

  • Kim, I.A.;Jin, E.J.;Cho, E.J.;Sohn, S.H.;Lee, C.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.563-570
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    • 2004
  • The insulin-like growth factor(IGF) system, consisting of IGFs-I and -II ligands and their receptors and six IGF-binding proteins(IGFBPs), plays an important role in survival, proliferation and differentiation of a variety of cell types. Lithium is a known modulator of survival and proliferation of many cell types in vitro. The present study was undertaken to investigate the relationship between LiCI-induced changes in cell survival and growth and the expression of the IGF system components in C6 rat glioma cell line which, besides IGF-I and its receptor, is known to express IGFBP-3 as its major IGF carrier. When C6 cells were cultured for 24h in the absence or presence of 2mM or 5mM LiCl in a 10% serwn-containing medium, the viability and the number of cells were not affected by added lithium. In 72-h culture, however, C6 cells clearly exhibited a dose-dependent response to added LiCl. The cells cultured for 72h in the presence of 0, 2mM and 5mM LiCl exhibited a typical mitotic, a growth-arrested and an apoptotic appearances, respectively. Moreover, the apoptotic cells were accompanied by reduced expression of IGF-I, IGF-I receptor and IGFBP-3 as examined by semi-quantitative reverse transcription-polymerase chain reaction. Interestingly, blockade of IGFBP-3 mRNA translation by addition of 101${\mu}M$ IGFBP-3 anti-sense oligodeoxyribonucleotide in serum-free, 24-h culture resulted in a decrease in the number of cells as well as relative abundance of the target mRNA. In summary, results suggest that the cytotoxic effect of lithium in C6 cell is likely to be mediated, in part, by suppression by this agent of the expression of the IGF system components. In this regard, IGFBP-3 may play at least a 'permissive' role in normal proliferation of this cell.