Dissertations / Theses on the topic 'Oligodeoxyribonucleotides'

A new highly hydrophobic 5'-hydroxyl protecting group (Tbf-DMTr) has been designed for the purification of synthetic oligonucleotides. Tbf-DMTr-oligonucleotides are strongly retained on RP-HPLC allowing a facile separation from truncated sequences. Subsequently the group can be removed in acidic conditions. The fluorescent properties of TBf-DMTr enable easy detection. The synthesis and purification of long oligonucleotides (> 100-mer) has been undertaken. Additionally a new fluorescent label for oligonucleotides has been developed which enables detection of DNA probes to concentration down to 10-¹⁰ mol/l.

Dans le but de cibler des oligonucleotides antisens diriges contre des genes du virus de l'immunodeficience humaine (vih) vers les monocytes et les macrophages, nous avons realise des conjugues oligonucleotide-serum albumine mannosylee (man-bsa) ou 6-phosphomannosylee (m6p-bsa) specifiquement reconnus et endocytes par les lectines de ces cellules. Des oligomeres resistants (alpha t#1#2) vis-a-vis des degradations enzymatiques et des oligonucleotides 19 meres beta d(#5aagctttatgaggcttaa#3) antisens complementaires de la region (+64, +63) des ltrs du vih ont ete realises. Ces sequences ont du etre modifiees sur l'extremite 3 par une fonction thiol, permettant leur attachement sur les transporteurs glycosyles, et marquees en position 5 par la fluoresceine ou la tyramine radioiodee. Differents conjugues oligonucleotide-neoglycoproteine (man-sab et m6p-sab) ont ete obtenus par couplage via un pont disulfure de ces oligomeres sur les proteines. D'autre part, des oligomeres portant un residu biotine en position 3 ont ete prepares et complexes avec la streptavidine mannosylee. L'etude de l'internalisation des oligonucleotides libres ou conjugues a ete realisee en cytometrie en flux, microscopie confocale et fractionnement cellulaire. Les resultats montrent que les oligomeres transportes par la man-sab, la m6p-sab ou le streptavidine mannosylee sont reconnus par les lectines membranaires de cellules du type macrophage et internalises selon un processus d'endocytose mediee par ces recepteurs. Le ciblage des oligonucleotides par les transporteurs glycosyles permet d'augmenter d'un facteur 10 a 20 la quantite d'oligonucleotide internalisee par les cellules et conduit a une concentration intracellulaire en oligomere 5 a 6 fois superieure a celle du milieu exterieur. Des experiences de pharmacocinetique sur la souris montrent que les transporteurs mannosyles ou 6-phosphomannosyles assurent le transport des oligonucleotides preferentiellement vers le foie, la rate et les poumons qui sont des organes riches en macrophages

Alkylating agents are a diverse family of compounds whose toxic, mutagenic and carcinogenic effects in living organisms are due to their ability to damage DNA. Humans are exposed to these agents through lifestyle, diet, occupation and some forms of chemotherapy, but they are also formed endogenously. To protect against these adverse effects, a variety of DNA repair process have evolved. Among these is O6-methylguanine-DNA methyltransferase (MGMT), a damage reversal protein that repairs O6-alkylguanine (O6-alkG) adducts by capturing the alkyl group onto a cysteine residue within the protein in an autoinactivating mechanism. Human MGMT is known to be present in at least 12 polymorphic variant forms and numerous studies have partially characterised one or a few of these in relation to cancer susceptibility. The main objective in this thesis was to compare the substrate specificity of seven MGMT variant proteins, and also a putative size-variant of the normal MGMT, using short ODNs containing 12 different O6-alkylguanines (O6-alkGs). A variety of analytical methods was used in these studies, including radioisotope-based assays, enzyme-linked immunosorbent assays (ELISA), mass spectrometry (MS) and surface plasmon resonance (SPR). The most potent inactivators of all MGMTs were ODNs containing O6-BzG followed by O6-MeG and O6-CMG. For the other ODNs, the MGMT proteins could be classed in three groups: firstly Wt, and the F84, V143/R178 and F84/V143/R178 variants for which the potencies were O6-PrG > O6-PobG > O6-EtG; secondly R160 and F84/R160 for which O6-PrG > O6-EtG > O6- PobG and finally Q128 for which O6- PobG > O6-PrG > O6-EtG. Alkyl group transfer from six of the ODNs to the active site cysteine of Wt MGMT was demonstrated by MS. The Q128 variant was the most resistant to inactivation by all modified ODNs. The R160 and F84/R160 variants were more resistant than the other variants to inactivation by the O6-CMG and O6-PobG containing ODNs. It was also shown that ZnCl2 supplementation during Wt MGMT expression in E.coli significantly increased MGMT specific activity and decreased sensitivity to inactivation by the ODNs. In addition both 5-methylcytosine adjacent and opposite O6-MeG significantly affected the inactivation of Wt MGMT. Finally, it was confirmed that O6-CMG is, contrary to previous reports, a substrate for MGMT, and the inefficient repair of O6-CMG by the E.coli Ogt alkyltransferase protein was shown to be the basis of the error. Studies with human cells suggest that MGMT also repairs O6-CMG in vivo. The results suggest that MGMT variants repair different O6-alkGs at different rates and hence the consequence of alkylating agents exposure will depend not only on the nature of the exposure but also an individual’s own specific MGMT variant.

The innate immune system provides an essential first line of defense against infection. Innate immune cells detect pathogens through several classes of Pattern Recognition Receptors (PRR) allowing rapid response to a broad spectrum of infectious agents. Activated receptors initiate signaling cascades that lead to the production of cytokines, chemokines and type I interferons all of which are vital for controlling pathogen load and coordinating the adaptive immune response. Detection of nucleic acids by the innate immune system has emerged as a mechanism by which infection is recognized. Recognition of DNA is complex, influenced by sequence, structure, covalent modification and subcellular localization. Interestingly certain synthetic oligodeoxynucleotides comprised of the TTAGGG motif inhibit proinflammatory responses in a variety of disease models. These suppressive oligodeoxynucleotides (sup ODN) have been shown to directly block TLR9 signaling as well as prevent STAT1 and STAT4 phosphorylation. Recently AIM2 has been shown to engage ASC and assemble an inflammasome complex leading to the caspase-1-dependent maturation of IL-1β and IL-18. The AIM2 inflammasome is activated in response to cytosolic dsDNA and plays an important role in controlling replication of murine cytomegalovirus (MCMV). In the second chapter of this thesis, a novel role for the sup ODN A151 in inhibiting cytosolic nucleic acid sensing pathways is described. Treatment of dendritic cells and macrophages with the A151 abrogated type I IFN, TNF-α and ISG induction in response to cytosolic dsDNA. A151 also reduced INF-β and TNF-α induction in BMDC and BMDM responding to the herpesviruses HSV-1 and MCMV but had no effect on the responses to LPS or Sendai virus. In addition, A151 abrogated caspase-1-dependent IL-1β and IL-18 maturation in dendritic cells stimulated with dsDNA and MCMV. Although inhibition of interferon-inducing pathways and inflammasome assembly was dependent on backbone composition, sequence differentially affected these pathways. While A151 more potently suppressed the AIM2 inflammasome, a related construct C151, proved to be a more potent inhibitor of interferon induction. A151 suppressed inflammasome signaling by binding to AIM2 and competing with immune-stimulatory DNA. The interaction of A151 and AIM2 prevented recruitment of the adapter ASC and assembly of the macromolecular inflammasome complex. Collectively, these findings reveal a new route by which suppressive ODNs modulate the immune system and unveil novel applications for suppressive ODNs in the treatment of infectious and autoimmune diseases. The innate immune response to HSV-1 infection is critical for controlling early viral replication and coordinating the adaptive immune response. The cytokines IL-1β and IL-18 are important effector molecules in the innate response to HSV-1 in vivo. However, the PRRs responsible for the production and maturation of these cytokines have not been fully defined. In the third chapter of this thesis, The TLR2-MyD88 pathway is shown to be essential for the induction of pro-IL-1β transcription in dendritic cells and macrophages responding to HSV-1. The HSV-1 immediate-early protein ICP0 has previously been shown to block TLR2 responses and in keeping with this finding, ICP0 blocked pro-IL-1β expression. Following translation, pro-IL-1β exists as an inactive precursor that must be proteolytically cleaved by a multiprotein complex known as the inflammasome to yield its active form. Inflammasomes are composed of cytoplasmic receptors such as NLRP3 or AIM2, the adapter molecule ASC, and pro-caspase-1. In the present study we found that the NLRP3 inflammasome is important for maturation of IL-1β in macrophages and dendritic cells responding to HSV-1. In contrast the related NLRP12 protein controls IL-1β production in neutrophils. These data indicate that sensing of HSV-1 by TLR2 drives pro-IL-1β transcription and infection activates the inflammasome to mature this cytokine. Moreover, these studies reveal cell type-specific roles for NLRP3 and NLRP12 in inflammasome assembly.

The innate immune system provides an essential first line of defense against infection. Innate immune cells detect pathogens through several classes of Pattern Recognition Receptors (PRR) allowing rapid response to a broad spectrum of infectious agents. Activated receptors initiate signaling cascades that lead to the production of cytokines, chemokines and type I interferons all of which are vital for controlling pathogen load and coordinating the adaptive immune response. Detection of nucleic acids by the innate immune system has emerged as a mechanism by which infection is recognized. Recognition of DNA is complex, influenced by sequence, structure, covalent modification and subcellular localization. Interestingly certain synthetic oligodeoxynucleotides comprised of the TTAGGG motif inhibit proinflammatory responses in a variety of disease models. These suppressive oligodeoxynucleotides (sup ODN) have been shown to directly block TLR9 signaling as well as prevent STAT1 and STAT4 phosphorylation. Recently AIM2 has been shown to engage ASC and assemble an inflammasome complex leading to the caspase-1-dependent maturation of IL-1β and IL-18. The AIM2 inflammasome is activated in response to cytosolic dsDNA and plays an important role in controlling replication of murine cytomegalovirus (MCMV). In the second chapter of this thesis, a novel role for the sup ODN A151 in inhibiting cytosolic nucleic acid sensing pathways is described. Treatment of dendritic cells and macrophages with the A151 abrogated type I IFN, TNF-α and ISG induction in response to cytosolic dsDNA. A151 also reduced INF-β and TNF-α induction in BMDC and BMDM responding to the herpesviruses HSV-1 and MCMV but had no effect on the responses to LPS or Sendai virus. In addition, A151 abrogated caspase-1-dependent IL-1β and IL-18 maturation in dendritic cells stimulated with dsDNA and MCMV. Although inhibition of interferon-inducing pathways and inflammasome assembly was dependent on backbone composition, sequence differentially affected these pathways. While A151 more potently suppressed the AIM2 inflammasome, a related construct C151, proved to be a more potent inhibitor of interferon induction. A151 suppressed inflammasome signaling by binding to AIM2 and competing with immune-stimulatory DNA. The interaction of A151 and AIM2 prevented recruitment of the adapter ASC and assembly of the macromolecular inflammasome complex. Collectively, these findings reveal a new route by which suppressive ODNs modulate the immune system and unveil novel applications for suppressive ODNs in the treatment of infectious and autoimmune diseases. The innate immune response to HSV-1 infection is critical for controlling early viral replication and coordinating the adaptive immune response. The cytokines IL-1β and IL-18 are important effector molecules in the innate response to HSV-1 in vivo. However, the PRRs responsible for the production and maturation of these cytokines have not been fully defined. In the third chapter of this thesis, The TLR2-MyD88 pathway is shown to be essential for the induction of pro-IL-1β transcription in dendritic cells and macrophages responding to HSV-1. The HSV-1 immediate-early protein ICP0 has previously been shown to block TLR2 responses and in keeping with this finding, ICP0 blocked pro-IL-1β expression. Following translation, pro-IL-1β exists as an inactive precursor that must be proteolytically cleaved by a multiprotein complex known as the inflammasome to yield its active form. Inflammasomes are composed of cytoplasmic receptors such as NLRP3 or AIM2, the adapter molecule ASC, and pro-caspase-1. In the present study we found that the NLRP3 inflammasome is important for maturation of IL-1β in macrophages and dendritic cells responding to HSV-1. In contrast the related NLRP12 protein controls IL-1β production in neutrophils. These data indicate that sensing of HSV-1 by TLR2 drives pro-IL-1β transcription and infection activates the inflammasome to mature this cytokine. Moreover, these studies reveal cell type-specific roles for NLRP3 and NLRP12 in inflammasome assembly.

Em 1978, o trabalho realizado por Stephenson e Zamecnik demonstrou a capacidade de um oligonucleotídeo de impedir a expressão de uma proteína específica. Atualmente, duas tecnologias são mais utilizadas para este propósito: os oligodeoxiribonucleotídeos antisense e o RNA de interferência (siRNA), que se aproveitam da capacidade de anelação entre as fitas complementares. A maior diferença entre as duas técnicas é a maquinaria proteica recrutada, isso é, o complexo RISC atua no funcionamento do siRNA, e a protease RNase H atua na clivagem da fita de RNA quando hibridizada com DNA. Apesar da grande aplicabilidade destas tecnologias, tanto para doenças metabólicas quanto para canceres, o veículo de entrega e proteção dessas sequências é de fundamental importância, visto que a aplicação desses oligonucleotídeos livres está sujeita à rápida degradação e ineficiência. A modificação das bases é uma das estratégias para conferir maior estabilidade às sequências, porém estas tem sido relacionadas a um aumento da toxicidade. Nessa dissertação, a quitosana, um polissacarídeo catiônico é utilizado para síntese de nanopartículas e encapsulamento dos oligodeoxiribonucleotídeos antisense (ASO). Para isso, foram realizadas modificações na quitosana comercial como despolimerização, trimetilação ou conjugação com PEG, seguida da síntese das nanopartículas com a adição de tripolifosfato de sódio (TPP) pelo método de gelatinização ionotrópica. A estabilidade das nanopartículas foi medida em função do tempo, da variação de temperatura e da diferença de pH. Além disso, a toxicidade dessas nanopartículas foi analisada através da viabilidade celular em diferentes linhagens, NB-4, HepaRG, HTC e BHK-570. A expressão da proteína verde fluorescente (GFP) na célula NB-4 foi utilizada para avaliar a entrega do ASO desenhado, sendo sua fluorescência monitorada por microscopia confocal. Os resultados demonstram que as nanopartículas se mantiveram estáveis durante o período de tempo analisado, assim como com a temperatura variando de 22 a 45°C e em pH ácido. Cada linhagem celular respondeu de forma diferente ao tratamento com as nanopartículas sem ASO, sendo a linhagem saudável BHK-570 com a maior resistência. Ademais, todas as células apresentaram viabilidade reduzida quando tratadas com concentrações na ordem de 1011 nanopartículas/mL a base de quitosana trimetilada. A fluorescência das células NB-4 quando tratada com as nanopartículas com ASO diminuiu consideravelmente nas 18 primeiras horas, seguida de um aumento após 42 horas. Dessa forma, pode-se concluir que as nanopartículas de quitosana propostas nessa dissertação apresentaram uma excelente alternativa para a entrega de material genético, principalmente para o trato gastro-intestinal, devido à sua estabilidade em pH ácido.
The property of an oligonucleotide to interfere in the expression of a protein was observed in 1978 by Stephenson and Zamecnik. To perform such interference, there are today, two main techniques being explored: antisense oligodeoxyribonucleotides and interference RNA. In both cases, the particularity of their chemical structure is taken into account as soon as they can bind in a complementary manner to the messenger RNA and inhibit its translation. The great difference between these techniques is related to the proteases involved in the process, while for interference RNA the RISC machinery acts, for antisense oligodeoxyribonucleotides RNase H cleaves the RNA in the duplex DNA-RNA. Although these tools to edit the translation process are relevant to the treatment and even cure of metabolic disorders and cancers, it is still not effective when employed without a coating to protect the sequences before it reaches the destiny in vivo. Efforts have been made in developing modified bases to be more stable, but they show some toxicity. In this dissertation, chitosan, a natural cationic polyssacharide, is used to produce nanoparticles to protect the antisense oligodeoxyribonucleotide (ASO). For this reason, the commercial chitosan was modified, depolymerized, trimetilated or PEGlated and the nanoparticles were synthesized with sodium tripolyphosphate (TPP) by ionotropic gelation method. The stability along time, in different pHs and temperatures was assessed. The toxicity of nanoparticles without ASO was quantified by MTT tests in NB-4, HepaRG, HTC and BHK-570 cell lines. A green fluorescent protein (GFP) expressed by NB-4 cells was the target to evaluate the delivery efficiency of the ASO, and its fluorescence was measured by confocal microscopy. Results showed that nanoparticles were stable over time as well as in temperatures ranging from 22 to 45°C and in acidic pH. Each cell line responded in a different manner to the treatment, with the health cell BHK-570 showing higher resistance. Furthermore, all of them presented lower viability when treated with trimetilated chitosan nanoparticles in the highest concentrations (ca 1011 nanoparticles/mL). NB-4 cells presented a decrease in fluorescence in 18 hours of treatment followed by an increase after 42 hours. We conclude that chitosan nanoparticles are a good alternative to the delivery of genetic material even more in the gastro intestinal tract due to its great stability in acid pH values.

La conformation des oligonucleotides (m**(5)dc-dg)::(2), (m**(5)dcdg)::(3) et (br**(5)dc-dg)::(3) peut etre sous forme b et z. Les stabilites thermiques de ces 2 formes sont differentes la fusion de la forme b est monophasique, celle de la forme est biphasique. Une premiere transition a ete observe, refletant un remaniement conformationnel intramoleculaire. Ce remaniement intramoleculaire conduit de la forme z a une double helice gauche dont le caractere de nucleotidique de l'unite repetitive est atteint et ou l'empilement des bases est modifie. L'etude du mecanisme d'echange chimique des protons de la guanine montre que dans tous les tampons utilisees catalysent le transfert en accord avec la theorie de biomoted. Etude comparative des cinetiques d'echange des formes z a permis d'attribuer les 2 protons lents aux protons amino de la cytosine

De nouvelles protections des fonctions amines en synthese oligonucleotidique ont ete recherchees afin de remplacer les groupements classiques trop stables. Ont ete selectiones le groupement phenoxyacetyle pour la protection de la desoxy-2' adenosine et de ladesoxy-2' guanosine et le groupement isobutyryle pour la protection de la desoxy-2' cytidine. Leur labilite permet une deprotection de l'oligonucleotide en fin d'assemblage en 2 heures u 20**(o)c au lieu de 17 heures u 60**(o)c. Des oligonucleotides de 11 a 72 bases de long ont ete assembles selon les trois principales methodes: phosphotriester, phosphoramidite et hydrogenophosphonate. Les nouveaux groupements ont permis d'autre part l'incorporation dans un fragment d'adn synthetique de nucleosides modifies fragiles en milieu alcalin. La dihydro-5,6 thymine, un des produits majeurs forme lors de la radiolyse de l'adn, a ainsi ete inseree dans plusieurs oligonucleotides. Cette possibilite devrait faciliter la comprehension des phenomenes de reparation cellulaire enzymatique ou de cancerogenese

De nombreuses modifications des bases de l'adn peuvent etre generees par divers facteurs comme les agents oxydants ou cancerigenes, les rayonnements afin d'evaluer les consequences biologiques et physico-chimiques de ces dommages, il est necessaire de posseder des modeles plus complexes de ceux-ci qui peuvent etre obtenus par leur incorporation dans des oligonucleotides par voie chimique. Ce travail est consacre a la preparation de fragments d'adn contenant des derives de la 2'-desoxyuridine. Le premier volet de ce travail a consiste a preparer un synthon phosphoramidite de la 5-formyl-2'-desoxyuridine et a l'incorporer dans des oligonucleotides de synthese. La suite de ce travail concerne la mise en evidence et la caracterisation d'une nouvelle lesion, resultant de l'oxydation de la thymidine: la 5-carboxy-2'-desoxyuridine. Par ailleurs, la preparation d'oligonucleotides comportant cette lesion a ete realisee. La troisieme partie de ce travail correspond a l'incorporation d'un troisieme defaut dans des oligonucleotides: la 5,6-dihydro-2'-desoxyuridine (dhdu). Une etude plus complete sur les produits de degradation de cette lesion, susceptibles de se former en milieu alcalin, a ete ensuite menee avec le monomere et avec un trinucleotide, d(gdhdut). Dans une derniere partie, la recherche de conditions analytiques de separation de produits d'oxydation de la thymidine par electrophorese capillaire est presentee

Le reverse dot est une methode appliquee a la detection simultanee de plusieurs sequences cibles par hybridation avec des sondes immobilisees sur un support. Nous proposons pour cela deux systemes de fixation covalente: immobilisation d'oligonucleotides 4-hydroxyphenyles sur des feuilles de cellulose diazotee et couplage des oligonucleotides amines sur des membranes de nylon fonctionnalise par de la polyethyleneimine. Pour les deux systemes, la fixation est stable et la capacite d'immobilisation est comparable a celle des autres methodes decrites. La selectivite pour le groupe phenol est superieure a 90% pour la cellulose diazotee alors qu'une fixation parasite par les nucleobases de 30% est observee pour le nylon. Une optimisation du procede sur nylon peut etre facilement envisagee. Les deux systemes ont ete testes pour la detection par hybridation des sequences des papillomavirus de type 16 et 6/11 amplifies par pcr. Une amorce fixee sur la cellulose permet d'effectuer l'amplification et la detection sur le meme support. Des phosphoramidites nucleosidiques et non nucleosidiques (derives de l'hexane-1,2,6-triol) ont ete prepares pour la fonctionnalisation des oligonucleotides par un groupe 4-hydroxyphenyle. Ces oligonucleotides modifies peuvent aussi etre utilises pour le marquage avec de l'iode 125 de sondes de detection. Ils peuvent, de plus, etre phosphoryles enzymatiquement: cela met en evidence la reconnaissance par la polynucleotide kinase du phage t#4 du groupe hydroxyle primaire de residus non-nucleosidiques portes en 5' d'un oligonucleotide. Plusieurs composes modeles, chiraux et achiraux, bases sur des motifs 1,2 et 1,3 diols ont ete etudies comme substrats. Nous avons montres que la phosphorylation est stereoselective pour les residus 1,2 diols

Synthèse et étude par spectroscopies RMN, IR et DC d'oligodésoxyribonucléotides contenant un résidu sans base: coexistence des conformations b et z sur un même duplex. Elaboration d'une base universelle dans la série des nucléosides pyrimidiques. Synthèse d'oligodésoxynucléotides ancrés à un support solide; applications.

Dans le but d'elucider les mecanismes moleculaires de la regulation de l'expression des genes, une etude structurale et fonctionnelle detaillee de la sequence tata du promoteur precoce du virus sv40 est realisee. La mutagenese dirigee utilisant des oligodesoxynucleotides de synthese permet la localisation des domaines fonctionnels de deux elements tata situes dans la region d'origine de replication virale. Chacun des elements dirige independamment l'initiation precise et efficace de la transcription precoce in vivo a partir d'un groupe de sites definis

A new method for the synthesis of N1-(beta- D-2'-deoxyribosyl)-5-methyl-2-pyrimidinone ( 1) was developed. Fluorescent nucleoside 1 was synthesized from thymidine in 5 steps and 40% overall yield. The 3-O-phosphoramidite of 1 was synthesized and oligodeoxyribonucleotides (ODNs) containing 1 were prepared in good yields using standard, automated solid-phase methodology. The ODNs were purified by RP-HPLC or PAGE. The purity of ODNs was analyzed by CE. Fluorescence spectra of the ODNs were recorded. The fluorescence of 1 was substantially quenched when 1 was incorporated into ODNs, and was quenched further after the formation of a double-stranded DNA. The binding of RecA protein to an ODN containing 1 resulted in an increase in the emission fluorescence yield in the absence or in the presence of ATPgammaS. The fluorescence emission and anisotropy changes that accompany the formation of the complexes have enabled us to establish by titration that RecA protein binds to about 3 nucleotides of ODN.

碩士
國立中興大學
生物化學研究所
87
Abstract The cleavage of DNA by vanadium(V)-preoxo complex under photo-irradiation conditions at neutral pH has been demonstrated (e.g., Inorg. Chem. 1997,36,1276-1277). In this study, we report the binding interaction between a vanadium(V)-peroxo complex, [VO(O2)2(5-NH2phen)] (where 5-NH2phen = 5-amino-1,10-phenanthroline), to seven synthetic oligo-nucleotides, 5’-GC-GATACC-GC---(I), 5’-GC-GGTATC-GC---(II), 5’-d(A)15---(IV) 5’-d(T)10---(V), 5’-AA-CCCCCC-TT---(VI) 5’-TT-GGGGGG-AA---(VII) and their mixture I+II、VI+VII, as well as a self-complementary oligermer 5’-GC-GATATC-GC---(III). The sequence-directed binding of oligo-DNA to vanadium(V)-peroxo complex was established. The fluorescence emission spectroscopy was used for monitoring the complex formation between vanadium(V)-preoxo complexes and DNA oligermers. The substrate III contains the recognition sequence 5’-GATATC- but not in substrates I, II, IV, V, VI, VII. In this study, the fluorescence emission of this complex was found to be quenched by all the substrates, which give linear Stern-Volmer plots. For substrate (III)2 (Tm = 22 oC), static quenching was identified from the temperature-dependent quenching data. These results therefore indicate that a preferential binding interaction exists between the complex at the single-stranded and double-stranded form of III. In summary, evidence for a specific binding interaction between the [VO(O2)2(5-NH2phen)]- complex and an oligodeoxynucleotide (III) containing the sequence 5’-GATATC was found. In addition, we also used CD spectroscopy and the UV/Visible spectroscopy demonstrated the duplex DNA that make form sample I+II、(III)2 and VI+VII which is B from duplex structure and the melting temperature(Tm) is near the room temperature.