【摘要】 目的在构建好的肽核酸压电基因传感器检测系统上引入“RecA蛋白互补单链DNA探针”复合体作为新型生物信号放大系统,提高传感器的检测灵敏度。方法压电基因传感器阵列表面先固定上乙肝病毒(HBV)的bisPNA探针,加入HBV基因组DNA与之反应完全后,加入“RecA蛋白互补单链DNA探针”与bisPNA/dsDNA复合物反应。首先分别优化RecA蛋白及ATPγS的浓度,再观察最佳条件下传感器检测系统的频率下降值和反应时间。结果当RecA蛋白浓度为3mg/ml,ATPγS浓度为12.5μmol/L时,所引起的频率下降值最明显。最佳条件下所引起的频率改变为(112.2±12.7)Hz。结论在检测系统中加入“RecA蛋白互补单链DNA探针”复合体,可有效地提高压电基因传感器的检测灵敏度。更多还原

【Abstract】 Objective To improve the detection sensitivity of peptide nucleic acid (PNA) piezoelectric gene sensor by introducing “RecA protein-complementary single strand DNA probe” complex as new style biological signal amplification system.Methods Bis-PNA probe for detecting HBV was immobilized on the surface of gene sensor array firstly.“RecA protein-complementary single strand DNA probe” was added to react with bis-PNA/dsDNA complex after bis-PNA probe was hybridized with corresponding target DNA completely.The concentrations of RecA protein and ATPγS were optimized respectively.And the frequency shift and reaction time were observed under the optimum conditions. Results The frequency shift arrived maximum when the concentration of RecA protein was 3.0 mg/ml and that of ATPγS was 12.5 μmol/L.The value of frequency shift was (112.2±12.7)Hz.Conclusion The sensitivity of piezoelectric gene sensor is improved effectively by adding “RecA protein-complementary single strand DNA probe” complex to the PNA detection system.更多还原