Teramoto N, Imanishi Y, Ito Y
Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, 606-8501, Japan.
Bioconjug Chem. 2000 Nov-Dec;11(6):744-8. doi: 10.1021/bc990146r.
A novel ligase ribozyme was in vitro selected from a random-sequence RNA library including N(6)-aminohexyl-modified adenine residues in place of natural adenine residues. This ribozyme mediated the formation of a phosphodiester bond with a DNA oligonucleotide through condensation with a 5'-triphosphate moiety on the ribozyme. Among the clones isolated from this selection, one was shown to accelerate ligation about 250-fold compared to the original random-sequence RNA library. Almost no rate acceleration was observed when the N(6)-aminohexyl-groups on adenine residues were omitted. Furthermore, ligation was dependent on the presence of a template DNA oligonucleotide that bridged the two strands.
一种新型连接酶核酶是从一个随机序列RNA文库中体外筛选出来的，该文库中用N(6)-氨基己基修饰的腺嘌呤残基取代了天然腺嘌呤残基。这种核酶通过与核酶上的5'-三磷酸部分缩合，介导了与DNA寡核苷酸形成磷酸二酯键。从该筛选中分离出的克隆中，有一个与原始随机序列RNA文库相比，连接速度加快了约250倍。当腺嘌呤残基上的N(6)-氨基己基基团被省略时，几乎没有观察到速度加快。此外，连接依赖于连接两条链的模板DNA寡核苷酸的存在。
