Compatibility of ten frequently used fluorescent proteins with EVOS Floid microscope
| Autor: | Stošić, Irin Antun |
|---|---|
| Přispěvatelé: | Svetec, Ivan Krešimir |
| Jazyk: | chorvatština |
| Rok vydání: | 2017 |
| Předmět: |
fluorescentna mikroskopija
fluorescentni proteini BIOTEHNIČKE ZNANOSTI. Biotehnologija Escherichia coli svjetleće diode fluorescent protein restriction analysis restrikcijska analiza Escherichia coli fluorescentna mikroskopija fluorescentni proteini restrikcijska analiza svjetleće diode fluorescence microscopy BIOTECHNICAL SCIENCES. Biotechnology light emitting diode |
| Popis: | Cilj ovog rada bio je istražiti kompatibilnost deset često korištenih fluorescentnih proteina s fluorescencijskim mikroskopom EVOS Floid, odnosno koji su od 10 proteina (mTagBFP2, mTFP1, mEGFP, mCitrine, tdTomato, mTagRFP, mCherry, mKate, mPlum i E2Crimson) vidljivi te mogu li se međusobno razlikovati ovim mikroskopom koji kao izvor svjetlosti koristi plave, crvene i zelene svjetleće diode. Plazmidi koji nose gene za fluorescentne proteine nabavljeni su u bakteriji E. coli, soj DH5α, izolirani su iz tog soja i njima je transformiran soj E. coli NEB Stable prikladniji za ekspresiju gena pod kontrolom promotora operona araBAD. Struktura plazmida potvrđena je restrikcijskom analizom, a mikroskopiranjem stanica E. coli s eksprimiranim fluorescentnim proteinima utvrđeno je da su EVOS Floid mikroskopom vidljivi svi proteini osim mTagRFP te da je u istom uzorku međusobno moguće razlikovati do pet različitih fluorescentnih proteina. The aim of this paper was to determine the compatibility of ten commonly used fluorescent proteins with the EVOS Floid fluorescence microscope. More specifically to determine which of the ten proteins (mTagBFP2, mTFP1, mEGFP, mCitrine, tdTomato, mTagRFP, mCherry, mKate, mPlum, and E2Crimson) are visible, and the way they differ from one another under this microscope that uses blue, red and green light emitting diodes as a light source. Plasmids carrying fluorescent protein genes were obtained in the DH5α E. coli strain, then isolated from this strain and the E. coli NEB Stable strain was transformed with them. The NEB Stable strain is more suitable for gene expression controlled by the araBAD promoter. The structure of plasmids was confirmed with restriction analysis. All proteins except for mTagRFP are visible with EVOS Floid microscope, and it is possible to distinguish between up to five fluorescent proteins in the same sample. |
| Databáze: | OpenAIRE |
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