Generation and characterization of an immortal cell line of xeroderma pigmentosum group E |
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| Affiliation: | 1. Department of Materials Science and Engineering, Fujian University of Technology, 350118 Fuzhou, China;2. Department of Materials Science and Engineering, The University of Tennessee, 37996–2200 Knoxville, TN, United States;3. Fujian Provincial Key Laboratory of Advanced Materials Processing and Application, 350118 Fuzhou, China;4. The Bredesen Center for Interdisciplinary Research and Graduate Education, The University of Tennessee, 37996–3394 Knoxville, TN, United States;1. Department of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 440-746, South Korea;2. Department of Marine Science, College of Natural Sciences, Incheon National University, Incheon 406-772, South Korea;3. Pacific Ocean Research Center, Korea Institute of Ocean Science and Technology, Ansan 426-744, South Korea;4. Department of Life Science, College of Natural Sciences, Hanyang University, Seoul 133-791, South Korea;5. Department of Marine Sciences and Convergent Technology, College of Science and Technology, Hanyang University, Ansan 426-791, South Korea;6. Department of Life Science, College of Natural Sciences, Sangmyung University, Seoul 110-743, South Korea;1. D. I. Mendeleyev University of Chemical Technology of Russia, Moscow, Russian Federation;2. Laboratory of Toxicology, University of Crete, Voutes, Heraklion, Crete, Greece;3. Department of Toxicology, University of Medicine and Pharmacy, Faculty of Pharmacy, Craiova, Romania;4. Department of Materials Science and Technology, University of Crete, University Campus Voutes, Heraklion, Crete, Greece |
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| Abstract: | Xeroderma pigmentosum group E (XP-E) fibroblasts (XP95TO) were transformed with pSV3neo. Selection in medium containing G418 yielded 14 clones with extended life span. Following crisis, one clone was recovered that behaved in culture as an immortal cell line and was named XPET6/1. Expression of the SV40 large T antigen gene (Tag) and increased level of p53 were demonstrated by western analyses. Fingerprinting with 14 polymorphic microsatellite genetic markers confirmed that XPET6/1 originated from the parental strain XP95TO. XPET6/1 retained the sensitivity to killing by UV observed with the parental strain. Cell-free extracts from the immortal or the parental XP-E cells were deficient in excision, compared to extracts from HeLa or extracts from Tag-transformed XP variant fibroblasts. Complementation of XP-E extracts with XP-A, XP-D or XP-G extracts restored nucleotide excision activity to normal levels. XPET6/1 could prove a useful cell line for cloning of the XPE gene by functional complementation. |
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