Reliable and quantitative electrochemical platform for direct detection of CYP707A1 gene expression from wheat leaves

Abstract

Gene-expression analysis is increasingly important in biological research, measuring the level of a particular gene expressed within a cell or tissue can provide a lot of valuable information. In this paper, we present a simple electrochemical (EC) biosensor that is capable of detecting CYP707A1 gene expression. Briefly, the target CYP707A1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) extraction from the leaves of Triticum aestivum (wheat). Due to the 5′ end of primers for PCR amplification modified with biotin and thiol separately, signal tag (Thi-dsDNA@SA-MB bioconjugate) was formed. The sensing platform was simply constructed by the assembly of typical gold nanostar and graphene with good repeatability. When the Thi-dsDNA@SA-MB bioconjugate was captured onto the sensing interface, numerous redox molecule thionine (Thi) could be introduced, producing a significant EC response. As a result, the proposed EC biosensor sensitively detects CYP707A1 express products of T. aestivum with the detection limit is as low as 13 pM. Importantly, the accuracy and reproducibility of this methodology, is comparable favorably with that of real-time RT-PCR (qRT-PCR), the benchmark technology for expression profiling of selected CYP707A1 gene in different drought-treated samples of T. aestivum.