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

recently been isolated, adapted to use prevalent human codons, and developed as a reporter for mammalian expression vectors by Stratragene.1 In contrast to the jellyfish Aequoria victoria enhanced GFP (EGFP), humanized Renilla GFP (hrGFP) has low cellular toxicity. This important attribute resolves problems that arise from the high cytotoxicity of EGFP, which triggers events that change the gene expression profile of cells and can confine experiments to the limited window of cell viability. In addition to maintaining cellular viability, hrGFP generates high-level fluorescence that can easily be detected in vivo by fluorescence microscopy or fluorescence activated cell-sorting (FACS) analysis.

Stratagenes Vitality vectors are specialized mammalian expression vectors that contain the hrGFP reporter. All Vitality vectors provide the following capabilities: generating stable cell lines expressing hrGFP as the reporter; detecting transfected cells via expression of hrGFP reporter; exchanging hygromycin, neomycin, and puromycin drug-resistance genes by Cre-Lox recombination; confirming transfection efficiency, and detecting fluorescence in vivo with the hrGFP reporter. Moreover, Vitality vectors are versatilethey are designed so that a gene of interest can be conveniently inserted in a variety of vectors for protein expression. Vitality vectors offer a variety of expression configurations that can be customized and optimized to best fit specific applications.

Bicistronic Expression Vectors

Fig.1

The pIRES-hrGFP-1a and pIRES-hrGFP-2a expression vectors (Figure 1) have a multiple cloning
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