The Green Fluorescent Protein Fluorophore HBDI And Its Mutant Molecules:Modification, Characterization And Properties

Green fluorescent protein (GFP) is a kind of excellent luminescence proteins, the modified synthesis of HBDI, which is the core fluorophore structure of GFP, is the hot topic in recent years. HBDI is often used as fluorescence labeling molecules, but rarely applied as fluorescent probe. This paper mainly relates to the synthesis of a series of modified HBDI molecules, the preparation of similar series of the HBDI molecules, (4-benzylidene-l-((2-hydroxynaphthalen-1-yl) methyl eneamino)-2-phenyl-lH-imidazol-5(4H)-one), the studies on their fluorescence properties, and the application as viscosity fluorescence probes as well. Finally a new synthesis method of 3-benzamide coumarin was found and its recognition of Fe3+ ions also studied. The main contents as follows:1. In the chapter 1, the research background of this thesis was described mainly, the composition of the fluorescent probe, the recognition mechanisms of fluorescent probe, and the research status of the fluorescent probe of GFP, HBDI and Schiffs base were emphasized.2. In the chapter 2, it mainly represented the synthesis and characterization of a series of modified HBDI mutant (4-benzylidene-1-((2-hydroxynaphthalen-1-yl)methyleneamino)-2- phenyl-1H-imidazol-5(4H)-one). Due to the lack of hydroxyl group which can bond to imidazoline ketone ring through hydrogen bond, therefore their fluorescence properties were not ideal, the continuing work remains to be completed.3. In the third chapter, a series mutant of the HBDI were designed and prepared (Compound 1, 2,3), and the corresponding fluorescence properties taking them as viscosity probe also studied. The results showed that there exist a good liner relation between log(fluorescence intensity) and log(viscosity).4. In the fourth chapter, a new synthesis method of 3-benzamide coumarin was found and its recognition to Fe3+ ions was studied. Compared with the traditional method, it has several advantages, such as, low difficulty, simple steps, economic raw materials. Furthermore, 3-benzamide coumarin indicates a good recognition to Fe3+ ion in ethanol solution, and shows good response to Fe3+ within the scope of 1-100 eq.5. In the fifth chapter some conclusions obtained in this paper were analyzed, and some prospects on following work were provided.