Murakami A, Tamura Y, Takeguchi S, Miyoshi H, Morii T, Maekawa T, Abe T, Makino K
Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
Nucleic Acids Symp Ser. 1993(29):21-2.
To investigate the mechanism of antisense effects, diastereoisomerism of oligodeoxynucleotides with modified internucleotide phosphate linkages such as oligodeoxynucleoside phosphoramidates (OPA) and oligodeoxynucleoside phosphorothioates (OPT). Reversed-phase HPLC was used to separate the diastereoisomers of OPA (up to tetramer) and OPT (up to pentamer), and it was found that the complete purification of all diastereoisomers could be achieved by the two-step chromatographic approach. From the circular dichroism measurements, it was found that some diastereoisomers formed certain hybrids with their complementary oligonucleotides and some did not.
为了研究反义效应的机制，研究了具有修饰的核苷酸间磷酸键的寡脱氧核苷酸的非对映异构现象，如寡脱氧核苷亚磷酰胺酯（OPA）和寡脱氧核苷硫代磷酸酯（OPT）。采用反相高效液相色谱法分离OPA（四聚体以下）和OPT（五聚体以下）的非对映异构体，发现通过两步色谱法可实现所有非对映异构体的完全纯化。通过圆二色性测量发现，一些非对映异构体与其互补寡核苷酸形成了特定的杂交体，而一些则没有。
