A single-label phenylpyrrolocytidine provides a molecular beacon-like response reporting HIV-1 RT RNase H activity.
A single-label phenylpyrrolocytidine provides a molecular beacon-like response reporting HIV-1 RT RNase H activity.
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DOI：
10.1093/nar/gkp1022
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发表时间：
2010-01
影响因子：
14.9
通讯作者：
Hudson RH
中科院分区：
文献类型：
--
作者：
Wahba AS;Esmaeili A;Damha MJ;Hudson RH
6-苯基吡咯胞苷（PhpC）是一种结构保守的高荧光胞苷类似物，被掺入到寡核糖核苷酸中。含有PhpC的RNA与互补DNA或RNA形成天然样双链体结构。发现PhpC修饰充当敏感的报告基团，对RNase H的结构和酶活性无破坏性。具有单个PhpC插入物的RNA/DNA杂合物是HIV-1 RT核糖核酸酶H的优良底物，并迅速报道了RNA链的切割，荧光强度增加了14倍。基于PhpC的RNase H测定在响应性、快速性和简易性方面上级传统的分子信标方法（单标记与双标记）。此外，基于PhpC的测定适合于高通量微孔板测定形式，并且可以形成用于HIV-RT RNA酶H抑制剂的新筛选的基础。
6-Phenylpyrrolocytidine (PhpC), a structurally conservative and highly fluorescent cytidine analog, was incorporated into oligoribonucleotides. The PhpC-containing RNA formed native-like duplex structures with complementary DNA or RNA. The PhpC-modification was found to act as a sensitive reporter group being non-disruptive to structure and the enzymatic activity of RNase H. A RNA/DNA hybrid possessing a single PhpC insert was an excellent substrate for HIV-1 RT Ribonuclease H and rapidly reported cleavage of the RNA strand with a 14-fold increase in fluorescence intensity. The PhpC-based assay for RNase H was superior to the traditional molecular beacon approach in terms of responsiveness, rapidity and ease (single label versus dual). Furthermore, the PhpC-based assay is amenable to high-throughput microplate assay format and may form the basis for a new screen for inhibitors of HIV-RT RNase H.
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影响因子：
14.9
作者：
Sandin P;Stengel G;Ljungdahl T;Börjesson K;Macao B;Wilhelmsson LM
通讯作者：
Wilhelmsson LM
影响因子：
56.9
作者：
COFFIN, JM
通讯作者：
COFFIN, JM
影响因子：
14.9
作者：
Dash, C;Rausch, JW;Le Grice, SFJ
通讯作者：
Le Grice, SFJ
影响因子：
16.6
作者：
Socher, Elke;Bethge, Lucas;Seitz, Oliver
通讯作者：
Seitz, Oliver
影响因子：
2.9
作者：
Hardman, Samantha J. O.;Thompson, Katherine C.
通讯作者：
Thompson, Katherine C.