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- W11386746 abstract "The identification and cloning of genes in the absence of knowledge of their corresponding proteins presents a challenge that may be overcome using expression vectors that complement a given phenotype. This approach involves the transfection of the cDNA of cells expressing a selectable phenotype into cells which lack this phenotype. The methodology requires a very high transfection efficiency, low background of spontaneous acquisition of the phenotype and is ultimately limited to identification of dominant genes. In this study we have sought to test the utility of an extrachromosomal-based host-vector system to identify the gene which confers the radiation sensitive phenotype to immortalized ataxia telangiectasia (AT) fibroblasts (1,2,3,4). We have adopted a protocol which allows for extrachromosomal maintenance of plasmids in host cells to effectively retrieve genes after transfection. This protocol is based on the phenomenon that in human or primate cell lines expressing the Epstein Barr Virus Nuclear Antigen 1 (EBNA1), plasmids that contain the Epstein Barr virus (EBV) origin of replication (Ori P) sequences will not integrate into the genome of host cells and will be maintained episomally (5,6,7).KeywordscDNA LibraryEpstein Barr VirusAtaxia TelangiectasiaAtaxia TelangiectasiaHigh Transfection EfficiencyThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves." @default.
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- W11386746 date "1991-01-01" @default.
- W11386746 modified "2023-09-26" @default.
- W11386746 title "Construction of a Unidirectional cDNA Library from a Radioresistant Laryngeal Squamous Cell Carcinoma Cell Line in an Epstein Barr Virus Shuttle Vector" @default.
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- W11386746 doi "https://doi.org/10.1007/978-1-4612-0411-4_38" @default.
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