通过模板导向化学连接实现寡核苷酸环化

Dolinnaya N G, Blumenfeld M, Merenkova I N, Oretskaya T S, Krynetskaya N F, Ivanovskaya M G, Vasseur M, Shabarova Z A

GENSET, Paris, France.

Nucleic Acids Res. 1993 Nov 25;21(23):5403-7. doi: 10.1093/nar/21.23.5403.

An efficient method for producing the covalent closure of oligonucleotides on complementary templates by the action of BrCN was developed. A rational design of linear precursor oligonucleotides was studied, and the effect of factors such as oligonucleotide concentration and oligomer-template length ratio was evaluated. The efficiency of circularization was shown to correlate well with the secondary structure of the precursor oligomer (as predicted by a simple computer analysis), hairpin-like structures bearing free termini clearly favouring the circularization reaction. A novel idea, consisting of the incorporation of non-nucleotide insertions in the precursor oligomer (namely, 1,2-dideoxy-D-ribofuranose residues), may render this method universal and highly effective. An original set of assays was developed to confirm the circular structure of the covalently closed oligonucleotides.

开发了一种通过溴化氰作用在互补模板上实现寡核苷酸共价封闭的有效方法。研究了线性前体寡核苷酸的合理设计，并评估了寡核苷酸浓度和寡聚物-模板长度比等因素的影响。结果表明，环化效率与前体寡聚物的二级结构（通过简单的计算机分析预测）密切相关，带有自由末端的发夹状结构明显有利于环化反应。一个新的想法，即在前体寡聚物中引入非核苷酸插入物（即1,2-二脱氧-D-核糖呋喃糖残基），可能使该方法具有通用性和高效性。开发了一套原始的检测方法来确认共价封闭寡核苷酸的环状结构。