Synthesis of base-modified oligonucleotides and DNA by polymerase incorporations of modified nucleoside triphosphates (dNTPs) is summarized. The crosscoupling synthesis of modified dNTPs, methods of their enzymatic incorporations, as well as their applications in bioanalysis and chemical biology are discussed.
- MeSH
- biochemie metody trendy MeSH
- diagnostické techniky molekulární MeSH
- DNA-dependentní DNA-polymerasy * MeSH
- nukleové kyseliny, nukleosidy a nukleotidy * chemická syntéza MeSH
- pojmy organické chemie MeSH
- polymerázová řetězová reakce MeSH
- techniky syntetické chemie MeSH
- techniky syntézy na pevné fázi * MeSH
- Publikační typ
- práce podpořená grantem MeSH
A novel efficient two-step methodology for the construction of base-functionalized DNA is based on direct aqueous cross-coupling reactions of unprotected nucleoside triphosphates followed by polymerase incorporation. Preliminary applications of the modified DNA in electrochemical detection and bioanalysis are outlined.
Construction of functionalized nucleic acids (DNA or RNA) via polymerase incorporation of modified nucleoside triphosphates is reviewed and selected applications of the modified nucleic acids are highlighted. The classical multistep approach for the synthesis of modified NTPs by triphosphorylation of modified nucleosides is compared to the novel approach consisting of direct aqueous cross-coupling reactions of unprotected halogenated nucleoside triphosphates. The combination of cross-coupling of NTPs with polymerase incorporation gives an efficient and straightforward two-step synthesis of modified nucleic acids. Primer extension using biotinylated templates followed by separation using streptavidine-coated magnetic beads and DNA duplex denaturation is used for preparation of modified single stranded oligonucleotides. Examples of using this approach for electrochemical DNA labelling and bioanalytical applications are given.
24 s. : il. ; 21 cm
- MeSH
- organická chemie MeSH
- puriny chemická syntéza farmakologie MeSH
- Publikační typ
- vysokoškolské kvalifikační práce MeSH
- Konspekt
- Chemie. Mineralogické vědy
- NLK Obory
- chemie, klinická chemie
- biochemie
A series of purine l-ribonucleosides 2a-2i bearing diverse C-substituents (alkyl, aryl, hetaryl or hydroxymethyl) in the position 6 were prepared by Pd-catalyzed cross-coupling reactions of 6-chloro-9-(2,3,5-tri-O-acetyl-beta-l-ribofuranosyl)purine with the corresponding organometallics followed by deprotection. Unlike their d-ribonucleoside enantiomers that possess strong cytostatic and anti-HCV activity, the l-ribonucleosides were inactive except for 6-benzylpurine nucleoside 2h showing moderate anti-HCV effect in replicon assay. A triphosphate of 2h did not inhibit HCV RNA polymerase.
- MeSH
- antivirové látky farmakologie chemická syntéza chemie MeSH
- financování organizované MeSH
- HeLa buňky MeSH
- Hepacivirus účinky léků MeSH
- HL-60 buňky MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- molekulární struktura MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- protinádorové látky farmakologie chemická syntéza chemie MeSH
- purinové nukleosidy farmakologie chemická syntéza chemie MeSH
- ribonukleosidy farmakologie chemická syntéza chemie MeSH
- screeningové testy protinádorových léčiv MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
