Synthetic Research Of Profluorescent Probe Detecting The RNA Transcription In Cells

Because of the existence of superiority such as diversity detection method, high sensitivity, strong visual and high detection speed, fluorescence probe techniques have been widely applied. Especially with the development of biotechnology, fluorescence probe techniques have been playing a more and more important role in protein, nucleic acid and cell detection.A fluorescent probe for detecting RNA transcription in cells based on the click reaction has been designed and synthesized in this paper. Firstly,5-ethynyluridine (EU) and 3-azido coumarin has been synthesized respectively. Secondly,5-ethynyluridine(EU) was incorporated into newly transcribed RNA via Bio-technology. Finally, EU-labeled cellular RNA was detected quickly by using a copper (l)-catalyzed cycloaddition reaction with 3-azido coumarin, followed by microscopic imaging.5-ethynyluridine(EU) was synthesized from the raw materials of uridine, acetic anhydride,ICl and trimethylsilyl acetylene through esterification, chloride, sonogashira coupling and ester alcoholysis reactions with the yield of 65%.3-azido coumarin was synthesized from salicylaldehyde, N-acetyl-glycine and sodium azide by the reactions of Schiff alkaline hydrolysis, diazolization and azidation with the yield of 86%. Sonogashira coupling, ester alcoholysis, Schiff alkaline hydrolysis and diazotization reactions are the four particularly critical steps of the nine-step total synthesis. The process conditions of the four steps were optimized through the experiments of single factor analysis.Besides, the products structure have been characterized by NMR, mass spectrometry, infrared and other analytical tools, through spectrum analysis confirming that the compound synthesized are the designed objectives.