Coleman R S, Pires R M
Department of Chemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210-1185, USA.
Nucleic Acids Res. 1997 Dec 1;25(23):4771-7. doi: 10.1093/nar/25.23.4771.
Full details of the template-directed covalent cross-linking of duplex oligodeoxynucleotides are presented. 4-Thio-2'-deoxyuridine was incorporated synthetically into a 17mer oligodeoxynucleotide, and the thiocarbonyl group of the modified base was alkylated with a variety of alpha-bromoacetyl-derivatized diamines. Covalent cross-linking was initiated by annealing the electrophilic probe oligomers with their complementary sequences, where a dG base was targeted at the position complementary to the modified 4-thio-2'-deoxyuridine. The sequence selectivity of cross-link formation as a function of tether topology and rigidity was examined, and the thermal stability of the modified duplexes was measured by UV melting experiments.
本文详细介绍了双链寡脱氧核苷酸的模板导向共价交联。将4-硫代-2'-脱氧尿苷通过合成方法掺入到一个17聚体寡脱氧核苷酸中,修饰碱基的硫羰基用多种α-溴乙酰基衍生的二胺进行烷基化。通过将亲电探针寡聚物与其互补序列退火来引发共价交联,其中在与修饰的4-硫代-2'-脱氧尿苷互补的位置靶向一个dG碱基。研究了交联形成的序列选择性作为连接拓扑结构和刚性的函数,并通过紫外熔解实验测量了修饰双链体的热稳定性。