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使用T4 RNA连接酶对tRNA和寡脱氧核苷酸进行荧光标记。

Fluorescent labelling of tRNA and oligodeoxynucleotides using T4 RNA ligase.

作者信息

Cosstick R, McLaughlin L W, Eckstein F

出版信息

Nucleic Acids Res. 1984 Feb 24;12(4):1791-810. doi: 10.1093/nar/12.4.1791.

Abstract

3'-O-(5'-phosphoryldeoxycytidyl) phosphorothioate and fluorescent 3'-O-(5'-phosphoryldeoxycytidyl) S-bimane phosphorothioate can be ligated to tRNA by T4 RNA ligase. They are also efficient donors for the enzymatic ligation to oligodeoxynucleotides bearing a 3'-cytidine terminus. Cytidine 3',5'-bisphosphate is also a substrate for the ligation reaction with DNA restriction fragments with a 3'-terminate cytidylic acid residue. Oligo- and polynucleotides with a 3'-phosphorothioate group react readily with electrophiles as exemplified by the reaction with monobromobimane.

摘要

3'-O-(5'-磷酸脱氧胞苷基)硫代磷酸酯和荧光3'-O-(5'-磷酸脱氧胞苷基)S-双硫代氨基甲酸盐可通过T4 RNA连接酶连接到tRNA上。它们也是与带有3'-胞苷末端的寡脱氧核苷酸进行酶促连接的有效供体。胞苷3',5'-二磷酸也是与带有3'-末端胞苷酸残基的DNA限制性片段进行连接反应的底物。带有3'-硫代磷酸酯基团的寡核苷酸和多核苷酸很容易与亲电试剂反应,与单溴双硫代氨基甲酸盐的反应就是一个例子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d69/318621/3b04e9d965a5/nar00322-0044-a.jpg

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