Zakharenko A L, Khodyreva S N, Rechkunova N I, Safronov I V, Pyshnyi D V, Degtyarev S K, Lavrik O I
Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of Russian Academy of Sciences, Novosibirsk, 630090, Russia.
Biochemistry (Mosc). 1998 Aug;63(8):929-34.
Substrate properties of dCTP analogs N4-[2-(4-azidotetrafluorobenzoylamino)-ethyl]-2;-deoxycytidine-5; -triphosphate (FABdCTP), 5-[N-(4-azidotetrafluorobenzoyl)-3-amino-trans-propen-1-yl] -2;-deoxycytidine-5;-triphosphate (AlFABdCTP), and N4-[2-(2-nitro-5-azidobenzoylamino)-ethyl]-2;-deoxycytidine-5; -triphosphate (NABdCTP) were studied in the reaction of DNA synthesis catalyzed by DNA polymerase from the extremely thermophilic bacterium Thermus thermophilus B 35 (Tte DNA polymerase). The enzyme was photoaffinity labeled with the mentioned derivatives, NABdCTP being used for the first time. The photoreactive primers containing FABdCTP and AlFABdCTP were synthesized in situ by Tte DNA polymerase and used in the complementary addressed labeling of DNA template. The efficiency of DNA template labeling is shown to be a function of the structure of the photoactive group.
研究了N4-[2-(4-叠氮基四氟苯甲酰胺基)-乙基]-2'-脱氧胞苷-5'-三磷酸（FABdCTP）、5-[N-(4-叠氮基四氟苯甲酰基)-3-氨基反式丙烯-1-基]-2'-脱氧胞苷-5'-三磷酸（AlFABdCTP）和N4-[2-(2-硝基-5-叠氮基苯甲酰胺基)-乙基]-2'-脱氧胞苷-5'-三磷酸（NABdCTP）等dCTP类似物在嗜热栖热菌B 35（Tte DNA聚合酶）的DNA聚合酶催化的DNA合成反应中的底物特性。该酶用上述衍生物进行了光亲和标记，其中NABdCTP是首次使用。含有FABdCTP和AlFABdCTP的光反应性引物由Tte DNA聚合酶原位合成，并用于DNA模板的互补寻址标记。DNA模板标记效率显示为光活性基团结构的函数。
