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| Detection of CaMV35S Gene Sequence in Transgenic Crops Using an Electrochemical Biosensor Based on Carboxylated Polypyrrole－Hemin and Point-triggered Strand Displacement Reaction Signal Amplification |
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| DOI： |
| KeyWord:carboxylated polypyrrole hemin point-triggered strand displacement reaction mimic peroxidase transgene electrochemical genosensor |
| Author | Institution |
| ZHAI Ying-hui，XIA Zi-hao，ZHANG Hong-yan，YE Yong-kang，CAO Xiao-dong，ZHENG Hai-song，LI Yun-fei |
1. School of Food Science and Biological Engineering，Hefei University of Technology，Hefei ，China; 2. School of Bioscience and Engineering，North Minzu University，Yinchuan ，China; 3. Technology Center of Hefei Customs District，Hefei ，China |
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| Abstract: |
| In order to meet the rapidly growing demands for safety evaluation and management on the genetically modified organisms（GMOs），the research and application of efficient and reliable GMOs detection technology has become more and more important．A novel electrochemical genosensor was prepared for sensitive detection of GMO components by utilizing the enhanced mimic peroxidase activity of cPPy－hemin to H2O2 and signal amplification strategy of point-triggered strand displacement reaction（TSDR） in this paper．Carboxylated polypyrrole（cPPy） and hemin（cPPy－hemin） nanocomposites were synthesized by a one-step method，which was used as signal tags to immobilize double-stranded DNA to form DNA－cPPy－hemin signal nanotags．Using the -NH2 groups in DNA duplex structure composed of signal probe（Ps），template strand（Ts） and auxiliary strand（As），the prepared cPPy－hemin was biofunctionalized to form Ts/As/Ps－cPPy－hemin．In presence of target transgenic cauliflower mosaic virus 35S promoter（CaMV35S） and fuel strand（Fs），the TSDR was triggered by the target CaMV35S，the released Ps－cPPy－hemin was captured by the immobilized capture probes（Cs） on the surface of GCE through hybridization．The quantitative detection of the target CaMV35S was realized by recording the catalytic reduction signal of H2O2 by cPPy-hemin. Under the optimal conditions，the fabricated biosensor had a good linear range of 1.0 × 10-14 -1.0 × 10-9 mol/L，with a detection limit（S/N = 3） of 3.2 × 10-15 mol/L．The biosensor possessed good selectivity，reproducibility and stability，which could be used for the detection of the CaMV35S segments in total DNA extracts from genetically modified soybeans. |
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