Engineering terminal disulfide bonds into DNA.
This protocol presents a simple and general means of modifying nucleic acids with disulfide cross-links. These cross-links serve as powerful tools to probe the structure, dynamics, thermodynamics, folding, and function of DNA and RNA, much in the way that cystine cross-links have been used to study proteins. The chemistry described has been used to synthesize disulfide-cross-linked hairpins and duplexes, higher-order structures such as triplexes, non-ground-state conformations, and tRNAs. since the cross-links form quantitatively by mild air oxidation and do not purturb either secondary or tertiary structure, this modification should prove quite useful for the study of nucleic acids.
Glick
Gary D
GD
University of Michigan, Ann Arbor, Michigan, USA.
D001483:Base Sequence; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004247:DNA / Q000138:chemical synthesis / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D015342:DNA Probes / Q000378:metabolism; D004220:Disulfides / Q000138:chemical synthesis*; D005818:Genetic Engineering / Q000379:methods*; D008954:Models, Biological; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes / Q000138:chemical synthesis / Q000737:chemistry; D009705:Nucleosides / Q000737:chemistry; D012107:Research Design; D013816:Thermodynamics
D003432:Cross-Linking Reagents; D015342:DNA Probes; D004220:Disulfides; D009692:Nucleic Acid Heteroduplexes; D009705:Nucleosides; C051802:triplex DNA; D004247:DNA