Biofunction-assisted DNA detection through RNase H-enhanced 3′ processing of a premature tRNA probe in a wheat germ extract
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- 作者：Atsushi Ogawa ; ogawa.atsushi.mf@ehime-u.ac.jp" class="auth_mail" title="E-mail the corresponding author ; Junichiro Tabuchi ; Yasunori Doi ; Masashi Takamatsu
- 关键词：DNA detection ; tRNA processing ; RNase H ; Cell-free translation ; Wheat germ extract
- 刊名：Bioorganic & Medicinal Chemistry Letters
- 出版年：2016
- 出版时间：1 August 2016
- 年：2016
- 卷：26
- 期：15
- 页码：3658-3661
- 全文大小：829 K
文摘
We have developed a novel type of biofunction-assisted, signal-turn-on sensor for simply and homogenously detecting DNA. This sensor system is composed of two types of in vitro-transcribed label-free RNAs (a 3′ premature amber suppressor tRNA probe and an amber-mutated mRNA encoding a reporter protein), RNase H, and a wheat germ extract (WGE). A target DNA induces the 3′ end maturation of the tRNA probe, which is enhanced by RNase H and leads to the expression of a full-length reporter protein through amber suppression in WGE, while there is almost no expression without the target due to the inactivity of the premature probe. Therefore, the target can be readily detected with the activity of the translated reporter. The catalytic reuse of the target with the help of RNase H in addition to various bioprocesses in WGE enables this sensor system to exhibit relatively high selectivity and sensitivity.