A 'one-tube' synthesis of a selective fluorescence 'turn off/on' DNA probe based on a C-phycocyanin-graphene oxide (CPC-GO) bio composite

被引:11
作者
Ghosh, Tonmoy [1 ,3 ]
Mondal, Aniruddha [2 ,3 ,5 ]
Vyas, Apeksha [1 ,4 ]
Mishra, Sandhya [1 ,3 ]
机构
[1] CSIR Cent Salt & Marine Chem Res Inst, Appl Phycol & Biotechnol Div, Gijubhai Badheka Marg, Bhavnagar 364002, Gujarat, India
[2] CSIR Cent Salt & Marine Chem Res Inst, Inorgan Mat & Catalysis Div, Gijubhai Badheka Marg, Bhavnagar 364002, Gujarat, India
[3] Acad Sci & Innovat Res AcSIR, Ghaziabad 201002, Uttar Pradesh, India
[4] Devi Ahilya Vishwavidyalaya, Sch Life Sci, Indore 452001, Madhya Pradesh, India
[5] Tatung Univ, Energy Storage & Convers Technol Res Ctr, Chem Engn Dept, Lab 405, Taipei 104, Taiwan
关键词
C-phycocyanin; Cyanobacteria; DNA sensing; Graphene oxide; Fluorescence quenching; ADSORPTION; REDUCTION; ENZYMES; TRENDS;
D O I
10.1016/j.ijbiomac.2020.06.286
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A naturally fluorescent protein, C-phycocyanin (CPC), was used as a fluorophore to study the effect of graphene oxide (GO) as a quencher. The protein was purified using established procedures and titrated with increasing GO concentrations. UV-visible titration showed a minor effect on the phycocyanobilin absorbance but significant interactions with the amino acid backbone. Fluorescence titration showed notable CPC quenching upon increasing GO concentration to 30 mu g ml(-1); the corresponding fluorescence dropped by similar to 97%. A non-linear Stern Volmer curve showed that the fluorophores did not interact directly with the quencher. Powder X-ray diffraction studies showed that the bio-composite lost the crystalline arrangement of GO and became amorphous, akin to CPC. SEM analysis showed GO sheets enfolding a protein nucleus with an increase in oxygen after the interaction compared to CPC. A 20 min incubation of the bio-composite with various biomolecules including amino acids, sugars, polydispersed exopolysaccharides (EPS), other proteins and DNA showed that only DNA could recover the CPC fluorescence. The `turn on' effect of DNA was distinguishable even when all the other molecules were in the same sample matrix. These results showed that CPC GO could be a fluorescence `turn off/on' DNA probe. (C) 2020 Elsevier B.V. All rights reserved.
引用
收藏
页码:977 / 984
页数:8
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