AN ASSAY FOR EXPRESS SCREENING OF POTATO TRANSFORMANTS BY GFP FLUORESCENCE
Abstract
An express assay for primary screening of potato transformants by their GFP fluorescence intensities is developed. In comparison to the widely used methods of transgenic plant screening by real-time reverse transcription polymerase chain reaction (Real-Time RT-PCR) or Northern-blotting, the GFP fluorescence assay needs n o expensive reagents and takes less time. This approach may allow to carry out nondestructive screening of the primary transgenic regenerants which can be further grown and used. To prove this assay reliability, the expression of the modified GFP (hGFP) gene in the leaves of transgenic potato (cv. “Skoroplodny”) plants, determined by its mRNA accumulation, was compared to GFP fluorescence intensity in the micro-samples of aseptic plant leaves. The strong correlation between the results of these two methods is the evidence of positive dependence of GFP fluorescence intensity on the target mRNA content.
About the Authors
N. O. Yuorieva
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
Russian Federation
Russian Federation
A. S. Voronkov
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences;
State Humanitarian-Technological University
Russian Federation
Russian Federation
D. V. Tereshonok
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
Russian Federation
Russian Federation
E. S. Osipova
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
Russian Federation
Russian Federation
E. V. Platonova
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
Russian Federation
Russian Federation
D. V. Belyaev
K.A. Timiryazev Institute of Plant Physiology, Russian Academy of Sciences
Russian Federation
Russian Federation
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Review
For citations:
Yuorieva N.O., Voronkov A.S., Tereshonok D.V., Osipova E.S., Platonova E.V., Belyaev D.V. AN ASSAY FOR EXPRESS SCREENING OF POTATO TRANSFORMANTS BY GFP FLUORESCENCE. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2018;73(2):85-92. (In Russ.)
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