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Generation of fusion genes carrying drug resistance, green fluorescent protein, and herpes simplex virus thymidine kinase genes in a single cistron.

Authors
Oh, SC; Nam, SY; Kwon, HC; Kim, CM; Seo, JS; Seong, RH; Jang, YJ; Chung, YH; Chung, HY
Citation
Molecules and cells, 11(2):192-197, 2001
Journal Title
Molecules and cells
ISSN
1016-84780219-1032
Abstract
We generated new fusion genes carrying positive- and negative-selection markers, and a reporter gene in a single reading frame. The new genes were constructed by sequentially linking the coding sequences of drug-resistance genes (hygro, or puro), a green fluorescence protein (GFP) gene (gfp), and the thymidine kinase gene (tk). The new synthetic genes (hygro/gfp/tk and puro/ gfp/tk) were inserted into retroviral vectors to test their usefulness as selective markers and reporters. The genes were functional in a positive selection in the presence of hygromycin (hygro/gfp/tk) or puromycin (puro/gfp/ tk). In addition, cells expressing the new fusion genes were clearly identifiable by their green fluorescence emitted from GFP. At the same time, these cells were sensitive to a gancyclovir treatment, allowing efficient removal of the transduced cells. The presently described synthetic genes will be valuable tools in both gene therapy and basic gene transfer studies, where positive selection of the transduced cells, monitoring gene expression, and negative selection of the transduced cells are simultaneously required.
MeSH terms
Anti-Bacterial Agents/pharmacologyAntiviral Agents/pharmacologyArtificial Gene Fusion*Cell LineDNA, Recombinant/geneticsDNA, Recombinant/metabolismDrug Resistance/genetics*Flow CytometryGanciclovir/pharmacologyGene Transfer TechniquesGenes/genetics*Genes, ReporterGenetic Vectors*Green Fluorescent ProteinsHumansHygromycin B/pharmacologyImmunoblottingLuminescent Proteins/genetics*Puromycin/pharmacologyRecombinant Fusion Proteins/geneticsRecombinant Fusion Proteins/metabolismRetroviridae/geneticsRetroviridae/metabolismSimplexvirus/enzymologySimplexvirus/geneticsThymidine Kinase/genetics*
PMID
11355700
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
AJOU Authors
장, 영주
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