The Construction And Application For Nucleic Acid Probes Of FoxM1

Nucleic acid probe is amolecular that can interact specially with target molecular,it also can be detected in apathognostic way.It is a new research method which can bedesigned, synthesised and applied in molecular level.It plays an important role topromote the development of molecular biology and molecular medicine.In this study we demonstrated a simple and efficient method based on FoxM1,onemember of Fox family,of SELEX to rapidly collect aptamers,we constructed twoaptamers that bind to FoxM1with high affinity and specificity.What’s more, a newapproach was developed for detection DNAzyme cleavage product based onmolecular beacon.The main researches included in this dissertation are presented asfollowing:1．We developed a simple and efficient method of SELEX to protein.wetransformed the right plasmids of PGEX-4T-1-GST-FoxM1-220-335andPGEX-4T1-GST into Escherichia coli and purified GST-FoxM1-220-335fusionprotein and GST.we got four aptamers that bind to FoxM1after eight rounds ofSELEX.2．A series of experiments just as fluorescence detection, ElectrophoreticMobility Shift Assay (EMSA),co-immunoprecipitation (co-ip) proved that ouraptamers bind to FoxM1with high affinity and specificity.3．A new method was developed for detection GST-FoxM1-220-335based onaptamer4．In the appendix B,we reported a new approach for detection DNAzymecleavage product based on molecular beacon (MB). MB was employed to monitormimic HCV-RNA product. Detection limit of the assay is0.02nmol/L.