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Dergiler / Sakarya University Journal of Science / 2021 / Cilt: 25 Sayı: 5

Development of a Fluorescent Protein Based FRET Biosensor for Determination of Protease Activity

İbrahim İNCİR
Özlem KAPLAN
Sema BİLGİN
İsa GÖKÇE
Sayfa
1235–1244
Yayın Tarihi
2021-10-30
DOI
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⚡ TL;DR(1 Cümlede Hızlı Çıkarım)

The usability of the designed FP-FRET biosensor in the determination of protease enzyme activity was demonstrated.

📌 3 Maddede Bu Makale

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Amaç & Hipotez:Proteases are closely associated with many pathological conditions.
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Temel Bulgular & Veri:The plasmid encoding the mNeonGreen-mRuby3 fluorescent protein-based FRET biosensor was constructed.
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Sonuç & Katkı:The usability of the designed FP-FRET biosensor in the determination of protease enzyme activity was demonstrated.
🔬Tür:Modelleme & Sayısal Simülasyon
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:Recombinant DNA Technology · Fluorescent Proteins · Fluorescent Resonance Energy Transfer (FRET)

Abstract

Proteases are closely associated with many pathological conditions. Efficient detection of protease activity may be useful for diagnosis, prognosis, and the development of new therapeutic biomolecules. Fluorescent Resonance Energy Transfer (FRET) is defined as the non-radioactive energy transfer that occurs between two fluorophores. Fluorescent proteins are widely used in FRET biosensors because they can be genetically encoded and compatible with cells. Fluorescent Protein based FRET (FP-FRET) biosensors are used to monitor biological processes such as enzyme activity, intracellular ion concentration, conformational changes, protein-protein interactions. In this study, it was aimed to detect protease activity using an FP-FRET biosensor and TEV protease was chosen as a model enzyme. The plasmid encoding the mNeonGreen-mRuby3 fluorescent protein-based FRET biosensor was constructed. The gene of the designed FP-FRET biosensor was expressed in Escherichia coli DH5α cells using recombinant DNA techniques and purified using Ni-NTA affinity chromatography. As a result, the activity of the TEV protease enzyme was determined by emission measurements performed in the spectrofluorometer using the produced FP-FRET biosensor. The usability of the designed FP-FRET biosensor in the determination of protease enzyme activity was demonstrated.

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Bu Makaleye Atıf Yap

İNCİR, İ., KAPLAN, Ö., BİLGİN, S., GÖKÇE, İ. (2021). Development of a Fluorescent Protein Based FRET Biosensor for Determination of Protease Activity. Sakarya University Journal of Science, Cilt: 25 Sayı: 5, 1235–1244. https://ajindex.com/articles/941766

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Kaynakça (13)

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