|
|
Thermodynamic Stability of Hybrid Nucleic Acid Complexes: Microcalorimetric Study
Ottová, Pavla ; Štěpánek, Josef (advisor) ; Vaisocherová, Hana (referee)
This work deals with implementation of differential scanning kalorimetry (DSC) into research of potential oligonucleotide therapeutics. Capabilities and limits of the DSC device have been determined and optimal conditions for measurement of nucleic acids complexes specified. Four types of modified oligonucleotides with isopolar, non-isosteric internucleotide linkage, consisting in insertion of a methylene group into the phosphodiester linkage between the phosphorus and one of the two ester oxygens (the methylene group inserted or at 3' or at 5' side of the linkage, the modified linkages alternate with the natural ones or all linkages are modified) were investigated. We determined thermodynamics characteristics of hybrid duplexes composed of modified deoxythymidine pentadecamers and complementary RNA polymer chain, polyadenylic acid. The oligonucleotide with the methylene group at the 3' side, the all linkages of which are modified, does not form complexes with polyadenylic acid. Other oligonucleotides form complexes and their stability increase in succession (3' end, alternating) ~ (5' end, all) < natural (dT)15 < (5' end, alternating). High cooperativity of the oligonucleotide to the polymer binding has been found out for all complexes. RNA hairpin of HIV-1 TAR element and its R06 aptamer have been also...
|
guest ::
RSS feed.