Gish G, Eckstein F
Max-Planck-Institut fuer experimentelle Medizin, Abteilung Chemie, Goettingen, West Germany.
Science. 1988 Jun 10;240(4858):1520-2. doi: 10.1126/science.2453926.
The difference in reactivity between phosphate and phosphorothioate diesters is the basis of a chemical degradation scheme for the sequencing of DNA and RNA. The phosphorothioate groups are incorporated into the nucleic acid in four separate enzymatic reactions, with three of the natural nucleoside triphosphates and one alpha-thiotriphosphate in each reaction. Selective strand cleavage is achieved through alkylation to form the hydrolytically labile phosphorothioate triester. As an example, the sequence analysis is presented of M13 phage DNA and of RNA prepared by transcription with SP6 RNA polymerase.
磷酸二酯和硫代磷酸二酯之间反应活性的差异是DNA和RNA测序化学降解方案的基础。硫代磷酸酯基团通过四个独立的酶促反应掺入核酸中，每个反应中有三种天然核苷三磷酸和一种α-硫代三磷酸。通过烷基化形成水解不稳定的硫代磷酸三酯实现选择性链切割。例如，展示了M13噬菌体DNA和通过SP6 RNA聚合酶转录制备的RNA的序列分析。
