We are analyzing https://link.springer.com/article/10.1007/s00438-002-0751-9.

Title:
Renilla luciferase-Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals | Molecular Genetics and Genomics
Description:
Light-emitting reporter proteins play an increasing role in the study of gene expression in vitro and in vivo. Here we present a ruc-gfp fusion gene construct generated by fusing a cDNA for Renilla luciferase (ruc) in-frame with a cDNA encoding the
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Keywords {🔍}

gene, article, gfp, fusion, renilla, rucgfp, luciferase, protein, fluorescence, reporter, access, privacy, cookies, content, wang, cells, information, publish, search, imaging, expression, aequorea, emission, data, log, journal, research, genomics, live, animals, shabahang, construct, transformed, presence, coelenterazine, open, discover, springer, optional, analysis, personal, parties, policy, find, track, molecular, genetics, luciferaseaequorea, dual, realtime,

Topics {✒️}

month download article/chapter renilla luciferase-aequorea gfp c-terminal lysine residue ruc-gfp fusion protein ruc-gfp fusion gene real-time imaging exciting gfp fluorescence article molecular genetics single-cell level based fusion gene construct privacy choices/manage cookies renilla luciferase activity related subjects full article pdf gfp fluorescence green fluorescence live animals bearing european economic area scope submit manuscript intramolecular energy transfer genomic dna purified metaphase chromosomes confirmed animal experiments revealed high-throughput screening conditions privacy policy fusion protein dual reporter accepting optional cookies aequorea gfp renilla luciferase loma linda university gene construct transformed cells exhibited october 2002 volume 268 journal finder publish ruc-gfp fluorescence live animals check access instant access article log gene expression luciferase substrate information privacy policy personal data genomics aims 'humanized' gfp article cite books a

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         headline:Renilla luciferase-Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals
         description: Light-emitting reporter proteins play an increasing role in the study of gene expression in vitro and in vivo. Here we present a ruc-gfp fusion gene construct generated by fusing a cDNA for Renilla luciferase (ruc) in-frame with a cDNA encoding the 'humanized' GFP (gfp) from Aequorea. A plasmid containing the fusion gene construct was successfully transformed into, and expressed in, mammalian cells. The transformed cells exhibited both Renilla luciferase activity in the presence of coelenterazine and GFP fluorescence upon excitation with UV light. Spectrofluorometry of cells containing the Ruc-GFP fusion protein, in the absence of wavelengths capable of exciting GFP fluorescence but in the presence of the luciferase substrate, coelenterazine, showed an emission spectrum with two peaks at 475 nm and 508 nm. These two peaks correspond to the emission maximum of Renilla luciferase at 475 nm and that of GFP at 508 nm. The peak at 508 nm generated in the presence of coelenterazine alone (without UV excitation) is the result of intramolecular energy transfer from Renilla luciferase to Aequorea GFP. Southern analysis of genomic DNA purified from transformed Chinese hamster ovary (CHO) cells and fluorescence in situ hybridization (FISH) to metaphase chromosomes confirmed the integration of the ruc-gfp fusion gene on a single chromosome. The bifunctional Ruc-GFP fusion protein allows the detection of gene expression at the single-cell level based on green fluorescence, and in a group of cells based on luminescence emission. Furthermore, animal experiments revealed that light emission from the Ruc-GFP fusion protein can be detected externally in the organs or tissues of live animals bearing the gene construct.
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      headline:Renilla luciferase-Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals
      description: Light-emitting reporter proteins play an increasing role in the study of gene expression in vitro and in vivo. Here we present a ruc-gfp fusion gene construct generated by fusing a cDNA for Renilla luciferase (ruc) in-frame with a cDNA encoding the 'humanized' GFP (gfp) from Aequorea. A plasmid containing the fusion gene construct was successfully transformed into, and expressed in, mammalian cells. The transformed cells exhibited both Renilla luciferase activity in the presence of coelenterazine and GFP fluorescence upon excitation with UV light. Spectrofluorometry of cells containing the Ruc-GFP fusion protein, in the absence of wavelengths capable of exciting GFP fluorescence but in the presence of the luciferase substrate, coelenterazine, showed an emission spectrum with two peaks at 475 nm and 508 nm. These two peaks correspond to the emission maximum of Renilla luciferase at 475 nm and that of GFP at 508 nm. The peak at 508 nm generated in the presence of coelenterazine alone (without UV excitation) is the result of intramolecular energy transfer from Renilla luciferase to Aequorea GFP. Southern analysis of genomic DNA purified from transformed Chinese hamster ovary (CHO) cells and fluorescence in situ hybridization (FISH) to metaphase chromosomes confirmed the integration of the ruc-gfp fusion gene on a single chromosome. The bifunctional Ruc-GFP fusion protein allows the detection of gene expression at the single-cell level based on green fluorescence, and in a group of cells based on luminescence emission. Furthermore, animal experiments revealed that light emission from the Ruc-GFP fusion protein can be detected externally in the organs or tissues of live animals bearing the gene construct.
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         Renilla luciferase Aequorea GFP (Ruc-GFP) fusion protein Reporter gene Renilla luciferase Aequorea GFP Fluorescence in situ hybridization (FISH)
         Plant Genetics and Genomics
         Human Genetics
         Microbial Genetics and Genomics
         Animal Genetics and Genomics
         Biochemistry
         general
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