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miRNA-319a
SENSORS AND ACTUATORS B-CHEMICAL · 2017 · IF 5.401

Photoelectrochemical detection of miRNA-319a in rice leaf responding to phytohormones treatment based on CuO-CuWO4 and rolling circle amplification

Bingchen Li,Huanshun Yin,Yunlei Zhou,Minghui Wang,Jun Wang,Shiyun Ai

MicroRNAs play a crucial role in regulating plant growth, such as involving in the pathways of phytohormons signaling, which function in broad range of important aspects in many stages of plant development and evolution. To figure out the components part of microRNA in phytohormone signaling network, a sensitive photoelectrochemical biosensor was developed to detect miRNA-319a specifically using CuO-CuWO 4 as photoactive material and in situ generation electron donor technique. Taking the advantage of rolling circle amplification, nicking endonuclease triggered exponential amplification of target molecule, specific interaction between phosphate group and phos-tag and between biotin and avidin, the developed strategy can greatly amplify the detection signal only using trace amount of target miRNA-319a. The wide linear range was obtained from 1 fM to 0.1   nM and the detection limit was estimated to be 0.47 fM (S/N   =   3). The developed method also showed good detection specificity. The effect of five phytohormones on the expression level of miRNA-319a in rice leaves was also investigated. This will be a basal principle of investigating the signaling network of phytohormones.

研究领域: 生物化学 · 分子生物学 · 分析化学 · 植物科学
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