PNA-Dependent Gene Chemistry: Stable Coupling of Peptides and Oligonucleotides to Plasmid DNA

• DOI: 10.2144/00282rr01
• Publication Date: 2018-08-29T11:27:54Z
• Authors: O. Zelphati; X. Liang; C. Nguyen; S. Barlow; S. Sheng; Z. Shao; P.L. Felgner
• Keywords: Streptavidin; Maleimide; Peptide nucleic acid

Two approaches are described for stably conjugating peptides, proteins and oligonucleotides onto plasmid DNA. Both methods use a peptide nucleic acid (PNA) clamp, which binds irreversibly specifically to binding site cloned into the plasmid. The first approach uses biotin-conjugated PNA clamp that can be used introduce functional biotin groups streptavidin bind. Atomic force microscopy images of linearized show localized at predicted on DNA strand. Peptides containing free thiol were conjugated maleimide streptavidin, these conjugates bound biotin-PNA-labeled In this way, peptides could brought stable association with A second maleimide-conjugated clamp. Methods thiolated directly maleimide-PNA-DNA hybrid. This straightforward technology offers an easy without disturbing its transcriptional activity.