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• W2034692015 abstract "To improve conventional yeast one‐hybrid screening, we have developed an efficient mammalian one‐hybrid system that allows rapid isolation of complementary DNAs which are able to induce human p14 ARF tumor suppressor gene. A 1.5 kb promoter region of p14 ARF was fused to EGFP to generate ARF promoter‐EGFP reporter vector. This reporter plasmid was stably trans‐fected into NH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F‐1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP‐positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP‐positive cells were prepared and the cDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert‐cloning site. This system should be useful for efficient screening of expression cDNA libraries in mammalian cells to identify novel upstream regulators for specific genes by one‐hybrid interaction." @default.
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• W2034692015 date "2002-01-01" @default.
• W2034692015 modified "2023-09-26" @default.
• W2034692015 title "Green fluorescent protein‐reporter mammalian one‐hybrid system for identifying novel transcriptional modulators for human p 14ARFtumor suppressor gene" @default.
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• W2034692015 doi "https://doi.org/10.1080/12265071.2002.9647670" @default.
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