Green fluorescent protein TurboGFP

- Extra fast protein maturation
- Bright green fluorescence
- Proven suitability to generate stably transfected cell lines
- Efficient maturation at a wide range of temperatures

Performance and use

TurboGFP can be expressed and detected in a wide range of organisms including cold-blooded animals. Mammalian cells transiently transfected with TurboGFP expression vectors give bright fluorescent signals within 8-10 hrs after transfection. No cell toxic effects and visible protein aggregation are observed.

	TurboGFP expression in transiently transfected mammalian cells.

TurboGFP suitability to generate stably transfected cells has been proven by Marinpharm company. Variuos cell lines expressing TurboGFP are commercially available.

Despite its dimeric structure, TurboGFP is suitable for generation of fusions. However, we recommend that you use specially optimized protein localization TagFPs to select a reporter for such purposes.

TurboGFP expression in stably transfected mammalian cell lines. (A) Human cervix carcinoma HeLa cells with TurboGFP in cytoplasm; (B) C2C12 mouse myoblasts with TurboGFP in cytoplasm; (C) PC-12 rat phaeochromocytoma cells with TurboGFP in cytoplasm; (D) PC-12 rat phaeochromocytoma cells with TurboGFP in cytoplasm after the addition of nerve growth factor (cells differentiate irreversibly into neuron-like cells); (G) Walker 256 rat tumour cells with TurboGFP in cytoplasm; (H) M3-mouse melanoma with TurboGFP in cytoplasm; (I) 3T3 mouse fibroblast with TurboGFP in cytoplasm; (J) T24 human bladder carcinoma with TurboGFP in cytoplasm; (E) Chinese Hamster Ovary Cells CHO-K1 with TurboGFP in cytoplasm; (F) HeLa cells expressing TurboGFP-fibrillarin fusion; (K) HeLa cells expressing mitochondria-targeted TurboGFP; (L) T24 human bladder carcinoma expressing mitochondria-targeted TurboGFP. Images were kindly provided by Dr. Christian Petzelt (Marinpharm).

TurboGFP maturation kinetics: TurboGFP allows monitoring the activity from early promoters. It matures noticeably faster than EGFP and most other fluorescent proteins. This difference in performance is illustrated here using both in vitro analysis of TurboGFP and EGFP refolding and maturation kinetics and in vivo examination of the developing Xenopus embrios expressing either TurboGFP or EGFP.

In vitro comparison of TurboGFP refolding and maturation kinetics with that of other fluorescent proteins showed higher TurboGFP maturation rate.

		Comparison of EGFP (violet lines) and TurboGFP (green lines) refolding and maturation speed in vitro. Normalized fluorescence recovery plots are shown. (A) — refolding kinetics; (B) — chromophore maturation kinetics.

In vivo examination of developing Xenopus embryos microinjected with vectors comprising either TurboGFP or EGFP under the control of CMV promoter showed bright fluorescence of TurboGFP immediately after midblastula transition, when gene expression is activated. At the same time, EGFP was practically invisible at this developmental stage. This example clearly demonstrates that TurboGFP is a better tool to study transgenic expression in rapidly developing embryos at early stages.

		In vivo comparison of TurboGFP and EGFP maturation in developing Xenopus embryos Vectors expressing the respective fluorescent proteins under the control of CMV promoter were microinjected into animal poles of Xenopus embryos at the stage of two blastomeres. Living embryos were then photographed from the animal pole at the middle and late gastrula stages. Experimental data were presented by Dr. A. Zaraisky, Institute of Bioorganic Chemistry, RAS (Moscow, Russia).