FUNCTIONAL TRANSCRIPTION ELONGATION COMPLEXES FROM SYNTHETIC RNA-DNA BUBBLE DUPLEXES
FUNCTIONAL TRANSCRIPTION ELONGATION COMPLEXES FROM SYNTHETIC RNA-DNA BUBBLE DUPLEXES
复制标题
DOI：
10.1126/science.1280856
复制
发表时间：
1992-11-20
期刊：
影响因子：
56.9
通讯作者：
VONHIPPEL, PH
中科院分区：
文献类型：
--
作者：
DAUBE, SS;VONHIPPEL, PH
设计并组装了一个合成的RNA-DNA气泡双工结构，旨在模仿功能性转录延伸复合物的核酸框架。该结构由可变长度的双链DNA（模板链和非模板链）组成，其中包含一个内部非互补的DNA“泡”序列。与模板DNA链部分互补的RNA寡核苷酸的3‘端在DNA泡内杂交，形成具有非互补的5’端RNA尾巴的RNA-DNA双工。将大肠杆菌或T7 RNA聚合酶添加到该构建物中形成一个复合物，该复合物以模板导向和进程的方式从杂交RNA引物中高效合成RNA，并表现出正常启动子启动的转录延伸复合物的其他特征。其他这样的结构可以设计来检查转录系统的许多功能和调控特性。
A synthetic RNA-DNA bubble duplex construct intended to mimic the nucleic acid frame-work of a functional transcription elongation complex was designed and assembled. The construct consisted of a double-stranded DNA duplex of variable length (the template and nontemplate strands) containing an internal noncomplementary DNA "bubble" sequence. The 3' end of an RNA oligonucleotide that is partially complementary to the template DNA strand was hybridized within the DNA bubble to form an RNA-DNA duplex with a noncomplementary 5'-terminal RNA tail. The addition of either Escherichia coli or T7 RNA polymerase to this construct formed a complex that synthesized RNA with good efficiency from the hybridized RNA primer in a template-directed and processive manner, and displayed other features of a normal promoter-initiated transcription elongation complex. Other such constructs can be designed to examine many of the functional and regulatory properties of transcription systems.