Design And Synthesis Of Functional DNA Phosphoramidite Monomer And Its Application In Bioanalysis

Based on the design and synthesis of functional DNA phosphoramidite monomer,DNA can be modified by different chemical small molecule with different biofunction for biological study.Based on it,this thesis includes the following three parts:1.Design of acrydite phosphoramidite and its application in DNA hydrogelBased on the design and synthesis of methylpropenyl phosphoramidite monomer,we develop new DNA hydrogel for biosensing.This part work develops target-responsive DNA hydrogel for AFB1 and glucose biosensing,respectively.For AFB1 analysis,DNA hydrogel combines with V-Chip for distance-readout portable and highly sensitive detection of AFB1.For glucose bioassays,the hydrogel system provides a non-enzymatic and visual method for glucose detection.2.Design of ortho-phthaladehyde phosphoramidite and its application in bioconjugation of DNA and proteinBased on the design and synthesis of ortho-phthaladehyde phosphoramidite monomer,we develop a new method of bioconjugation between DNA and protein.As the rapid and high efficient reaction between OPA and primary amine condensation reaction with the formation of phthalimidines under mild condition,OPA-DNA is synthesized and applied in bionconjugation with protein.This work choose cytochrome c and lysozyme as model protein to investigate the coupling efficiency.The results demonstrate that OPA-DNA can react with cytochrome c and lysozyme quickly and efficiently.The OPA-DNA can also interact with NH2-cDNA in a fast and efficient coupling.This DNA probe has promising applications in the bioconjugation of DNA and protein.3.Design of tetra-ortho-methoxyazobenzene phosphoramidite and its application in photo regulation of DNA hybridizationBased on the design and synthesis of tetra-ortho-methoxyazobenzene phosphoramidite monomer,we develop a new DNA probe which can proceed photo regulation with visual light.Experimental results demenstrates the regulation effect in mild green light and blue light at the molecular level.Then,HPLC results characterize the regulation effect of Azo-DNA with visual light.UV-vis spectra characterizes the melting temperature change under different light,which indicates visual light can regulate DNA hybridization.The DNA probe is used in T7 RNA transcription system,which can realize T7 RNA transcription regulation with visual light.By modifying tetra-ortho-methoxyazobenzene,visual light can regulate the DNA sequence,which has promising applications in biological regulation systems.