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[用于通过非放射性方法研究核酸的核苷-5'-磷酸荧光类似物]

[Fluorescent analogs of nucleoside-5'-phosphates for the study of nucleic acids by nonradioactive methods].

作者信息

Aleksandrova L A, Lukin M A, Rozovskaia T A, Atrazhev A M, Kukhanova M K, Kraevskiĭ A A

出版信息

Mol Biol (Mosk). 1990 Jul-Aug;24(4):1100-8.

PMID:1701217
Abstract

The synthesis of 2'-deoxyuridine 5'-triphosphate analogues with fluorescent residues of fluorescein and rhodamine nature at C5 of the uracil base was performed. Reverse transcriptase of avian myeloblastosis virus, DNA polymerase beta of rat liver, terminal deoxynucleotidyl transferase of calf thymus and E. coli DNA polymerase I, Klenow fragment, were shown to be capable to incorporate a nucleotide residue with fluorescent label into 3'-terminus of oligonucleotide. These fluorescent labeled oligonucleotides were used as primers for synthesis of (-)-chain of M13mp10 phage. Fluorescently labeling template-primer complexes were used for DNA sequencing.

摘要

进行了在尿嘧啶碱基的C5位带有荧光素和罗丹明类荧光残基的2'-脱氧尿苷5'-三磷酸类似物的合成。结果表明,禽成髓细胞瘤病毒逆转录酶、大鼠肝脏DNA聚合酶β、小牛胸腺末端脱氧核苷酸转移酶和大肠杆菌DNA聚合酶I的Klenow片段能够将带有荧光标记的核苷酸残基掺入寡核苷酸的3'-末端。这些荧光标记的寡核苷酸被用作合成M13mp10噬菌体(-)链的引物。荧光标记的模板-引物复合物用于DNA测序。

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