Studies On Self-assembled Bilayer Lipid Membranes And Immobilization Technique Of Gene Biosensor

In this study, some techniques which have close relation to biosensor including the method to construct an bilayer lipid membranes (BLMs) with the tip of patch clamp pipette (patch pipette-BLMs) and the method to immobilize the DNA probes specific for Listeria monocytogenes(LM) on gold electrode of quartz crystal microbalance(QCM) were explored. The contents are as follows:1. Succeed in constructing the patch pipette-BLMs and its reality was verified by embedding gramicidin in it. This work is the foundation for the further study on the application of patch pipette-BLMs in ion channel and fast test of ultra-trace substance.2. DNA probes with good LM specificity and fit for constructing QCM-DNA sensor were designed. The DNA probes not only can be used for developing QCM-DNA sensor but also can be used in other tests for LM which by application of DNA hybridization or PCR techniques.3. A new activity couplet TAPD (N-[6-(thien-3-yl)acetoxyl]-pyrrolidine-2, 5 -dione) was synthesized in the study. TAPD can self-assembling on gold electrode, and the DNA probes linked by it would have better space for DNA hybridization. Otherwise. TAPD do not need to be activated in the coupling reaction. TAPD can also link other biomolecular such as antibody and enzyme to the gold electrode.4. Study of the method of immobilizing the DNA probes on QCM gold electrode through the couplet TAPD. The method can fix the DNA probes effectively and the procedure is simple, easy handling and time-saving, which have practical value. The preliminary test data showed the QCM-DNA sensor developed by this method can real-time detect the DNA hybridization. And this method can also be used in developing other type biosensor such as SPR.