Open AccessDesign and Application of γ-aminobutyric Acid Nano-Fluorescent Probe
Author(s) -
Qiao Hu,
Ying Pang,
Yuxin Shi,
Ke Yang,
Xiaoling Liao,
Xue Li
Publication year - 2020
Publication title -
iop conference series. materials science and engineering
Language(s) - English
Resource type - Journals
eISSN - 1757-899X
pISSN - 1757-8981
DOI - 10.1088/1757-899x/774/1/012109
Subject(s) - bacteria , agarose , green fluorescent protein , fluorescence , plasmid , subcloning , förster resonance energy transfer , escherichia coli , biochemistry , recombinant dna , biology , metabolite , chromatography , chemistry , biophysics , dna , genetics , gene , physics , quantum mechanics
The design and development of nano-fluorescent probes is a research hotspot in the field of biological functional materials. The construction of γ-aminobutyric acid (GABA) fluorescent probe based on fluorescence resonance energy transfer (FRET) technology will become a new tool for bacterial screening. Recombinant plasmids suitable for prokaryotic expression were constructed by subcloning technology, and verified by agarose gel electrophoresis and sequencing. The target protein was expressed in BL21 E. coli and purified by nickel matrix affinity chromatography. Spectroscopy and fluorescence imaging validate probe function and evaluate bacteria’s environmental adaptability. The results showed that a recombinant plasmid suitable for efficient and stable expression of prokaryotes was successfully constructed. The release of GABA will cause changes in fluorescence intensity. The probe can evaluate the metabolic activity and adaptability of bacteria to different environments. This method uses bacteria’s metabolic activity to realize the selection of bacteria. The sample does not require special biochemical treatment. Compared with traditional evaluation methods by counting the number of bacteria and analyzing metabolite, it is a more convenient tool for screening and evaluation of bacteria strains.