Biomedical subjects
Publications and source records attributed to Yuichiro Aiba.
By introducing an intercalator through D-threoninol to the 10-23 DNAzyme at the junction between its catalytic loop and the binding arm, the RNA cleavage activity was greatly improved.
Recently, we developed artificial restriction DNA cutter (ARCUT) composed of pseudo-complementary peptide nucleic acid (pcPNA) and Ce(IV)/EDTA complex (EDTA = ethylenediamine-N,N,N',N'-tetraacetate). Here we promoted the site-selective hydrolysis by attaching phosphate groups to the pcPNAs. The promotion by the phosphates increased with decreasing length of the gap-like site. Furthermore, the scission was successful even when phosphate groups were introduced to 0 base-gap system.
In order to hydrolyze double-stranded DNA efficiently at the target site, two pseudo-complementary peptide nucleic acids (pcPNAs) bearing phosphate group were combined with Ce(IV)/EDTA complex (EDTA = ethylenediamine-N,N,N',N'-tetraacetate). The phosphate groups as metal-binding ligands were placed near the target site, and concentrated the Ce(IV) complex thereto. As the result, the site-selective hydrolysis was notably promoted, compared with the scission by the cutters involving unmodified pcPNAs.
Artificial restriction DNA cutter (ARCUT), which we developed recently, was used for manipulation of plasmid DNA. PCR product was inserted into pBR322 plasmid vector using ARCUT, and E. coli cells were transfected with this recombinant plasmid DNA. Successful growth of cells shows that the recombination proceeds without any unexpected mutations. Furthermore, this artificial system was applied to PCR-free construction of chimera protein.