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New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence ,,, Humanized Renilla GFP Reporter

tor, we analyzed the fluorescence profile of transduced cells by FACS. These data showed that ~100% of the cells were transduced with pFB-hrGFP and that the fluorescence from hrGFP expression was vivid (data not shown).

Conclusions

The Vitality hrGFP vectors, which contain the humanized green fluorescent protein from Renilla reniformis (hrGFP), are useful mammalian expression vectors for monitoring gene expression and localizing proteins. Now, the hrGFP gene is available in a variety of vector formats for choosing the appropriate configuration for specific gene expression studies. In a variety of experiments, we used the original Vitality pIRES-hrGFP-1a and pIRES-hrGFP-2a vectors to express epitope-tagged proteins. We used the new Vitality phrGFP-N1 and phrGFP-C vectors to express hrGFP-protein fusions that targeted specific subcellular locations. In further tests, we used the Vitality pFB-hrGFP retroviral control vector to transduce a variety of mammalian cells. These tests confirmed the functionality and utility of these vectors and the high-intensity fluorescence of hrGFP expression. Most importantly, the low toxicity of hrGFP expression ensures that use of Vitality vectors results in artifact-free transient and stable experiments.

REFERENCES

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2. Fieck, A., et. al. (1995) Nucleic Acids Res. 20: 1785-91.
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5. Gould, SG., et.al.. (1997) J. Cell Biol
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