A Novel Fluorescent GFP Chromophore Analog-Based Dye for Quantitative PCR
- Authors: Stakheev A.A.1, Ryazantsev D.Y.1, Zvezdina Y.K.1, Baranov M.S.1, Zavriev S.K.1
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Affiliations:
- Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
- Issue: Vol 83, No 7 (2018)
- Pages: 855-860
- Section: Short Communications
- URL: https://journals.rcsi.science/0006-2979/article/view/151691
- DOI: https://doi.org/10.1134/S000629791807009X
- ID: 151691
Cite item
Abstract
This is the first report describing the possibility of using a green fluorescent protein chromophore synthetic analog, P-HOBDI-BF2, as a fluorescent dye for a linear hydrolysis probe used in qPCR. The study was carried out on a system for detection of the plant pathogenic fungus Fusarium avenaceum using a plasmid containing translation elongation factor 1α fragment as a template. To estimate fluorogenic properties of P-HOBDI-BF2, 6-FAM-and BDP-FL-labeled probes were used. It was demonstrated that a synthetic dye based on the P-HOBDI-BF2 chromophore can be used for labeling hydrolysis probes for qPCR, but fluorescence increase levels for P-HOBDI-BF2-labeled probes were slightly lower than those for 6-FAM-labeled ones. At the same time, the sensitivity of P-HOBDI-BF2-based assays remained high, and this fact together with acceptable fluorescence levels suggests that this dye can be considered as an efficient alternative for reporters traditionally used for fluorescence detection in the FAM channel.
About the authors
A. A. Stakheev
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
Author for correspondence.
Email: stakheev.aa@gmail.com
Russian Federation, Moscow, 117997
D. Yu. Ryazantsev
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
Email: stakheev.aa@gmail.com
Russian Federation, Moscow, 117997
Yu. K. Zvezdina
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
Email: stakheev.aa@gmail.com
Russian Federation, Moscow, 117997
M. S. Baranov
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
Email: stakheev.aa@gmail.com
Russian Federation, Moscow, 117997
S. K. Zavriev
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry
Email: stakheev.aa@gmail.com
Russian Federation, Moscow, 117997