Sawai Hiroaki, Wada Makoto, Kouda Tsukasa, Nakamura Ozaki Akiko
Department of Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Chembiochem. 2006 Apr;7(4):605-11. doi: 10.1002/cbic.200500291.
5'-pACUG tetraribonucleotides containing 2'-5' or 3'-5' linkages self-condensed on 2'-5'- or 3'-5'-linked complementary decaribonucleotide (5'-CAGUCAGUCA) templates. CD and UV melting studies showed that helix formation took place in all four possible combinations of linkage isomers of the substrate tetramer and the template decamer under the ligation conditions. The hybridization ability followed the order: [3'-5' tetramer with 3'-5' decamer] > [2'-5' tetramer with 3'-5' decamer] > [2'-5' tetramer with 2'-5' decamer] > or = [3'-5' tetramer with 2'-5' decamer]. Each tetramer condensed on the complementary decaribonucletide template to form the corresponding octamer, but the ligation efficiency varied considerably, depending on the types of linkage in the tetramer substrate and the template decamer. The yields of the octamers obtained by the template-directed ligation followed the order: [2'-5' substrate: 2'-5' template] > [3'-5':3'-5'] > [2'-5':3'-5'] > [3'-5':2'-5']. The results demonstrate that a homo-linkage system is preferable for the template-directed synthesis of RNA. The resulting linkage of the octamer formed from the 2'-5'-linked substrate and the 2'-5'-linked template is mainly 2'-5'.
含有2'-5'或3'-5'连接的5'-pACUG四核糖核苷酸在2'-5'-或3'-5'-连接的互补十脱氧核糖核苷酸(5'-CAGUCAGUCA)模板上进行自缩合。圆二色光谱(CD)和紫外熔解研究表明,在连接条件下,底物四聚体和模板十聚体的连接异构体的所有四种可能组合中均发生了螺旋形成。杂交能力遵循以下顺序:[3'-5'四聚体与3'-5'十聚体]>[2'-5'四聚体与3'-5'十聚体]>[2'-5'四聚体与2'-5'十聚体]≥[3'-5'四聚体与2'-5'十聚体]。每个四聚体在互补的十脱氧核糖核苷酸模板上缩合形成相应的八聚体,但连接效率差异很大,这取决于四聚体底物和模板十聚体中的连接类型。通过模板导向连接获得的八聚体产率遵循以下顺序:[2'-5'底物:2'-5'模板]>[3'-5':3'-5']>[2'-5':3'-5']>[3'-5':2'-5']。结果表明,对于RNA的模板导向合成,同型连接系统是更可取的。由2'-5'-连接的底物和2'-5'-连接的模板形成的八聚体的最终连接主要是2'-5'。