Doubly Catalytic Sensing of HIV-1-Related CCR5 Sequence in Prokaryotic Cell-Free Translation System using Riboregulator-Controlled Luciferase Activity
详细信息 查看全文
- 作者：Shinsuke Sando ; Atsushi Narita ; Kenji Abe ; and Yasuhiro Aoyama
- 刊名：Journal of the American Chemical Society
- 出版年：2005
- 出版时间：April 20, 2005
- 年：2005
- 卷：127
- 期：15
- 页码：5300 - 5301
- 全文大小：88K
- 年卷期：v.127,no.15(April 20, 2005)
- ISSN：1520-5126
文摘
A molecular-beacon-type riboregulator (mRNA) was applied to multiply catalytic gene sensing. It consists of a reporter gene for firefly protein luciferase and, upstream thereof, a regulator hairpin domain composed of an RBS/anti-RBS stem (RBS = ribosome binding site) and a loop which is complementary to the target. The hairpin and, hence, the RBS are rendered open upon binding of a target oligonucleotide of the human CC chemokine receptor 5 sequence in a prokaryotic cell-free translation system (10 
L) to ignite ribosomal catalytic translation, or transcription/translation when using a DNA form of the probe, to produce luciferase, which is assayed by a catalytic chemiluminescence reaction. The sensing, using an unmodified RNA or even dsDNA as a probe with a chemiluminescence output, is thus doubly catalytic or amplifiable with a sensitivity at 
50 fmol in respect to the target with 4.5 fmol (1 ng/L) of probe and a single nucleotide resolution.
L) to ignite ribosomal catalytic translation, or transcription/translation when using a DNA form of the probe, to produce luciferase, which is assayed by a catalytic chemiluminescence reaction. The sensing, using an unmodified RNA or even dsDNA as a probe with a chemiluminescence output, is thus doubly catalytic or amplifiable with a sensitivity at 50 fmol in respect to the target with 4.5 fmol (1 ng/L) of probe and a single nucleotide resolution.