1995年6月
A NEW RETROVIRAL VECTOR FOR DETECTING MUTATIONS AND CHROMOSOMAL INSTABILITY IN MAMMALIAN-CELLS
MUTATION RESEARCH-ENVIRONMENTAL MUTAGENESIS AND RELATED SUBJECTS
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- 巻
- 334
- 号
- 3
- 開始ページ
- 375
- 終了ページ
- 383
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1016/0165-1161(95)90075-6
- 出版者・発行元
- ELSEVIER SCIENCE BV
A retroviral vector carrying both forward (neo) and backward (herpes simplex virus thymidine kinase or HSV-TK gene) selection markers was constructed as a substrate for mutational assay in mammalian cells. The cells infected with this virus are first selected with G418, mutagenized and then selected with the anti-herpes drug acyclovir (ACV). Since HSV-TK, but not the host TK, is capable of converting ACV to a toxic metabolite, cells retaining the intact HSV-TK gene fail to survive, while the cells carrying a mutated HSV-TK gene or which have lost the gene can form colonies in the presence of ACV, making it possible to detect the genetic defects in a positive manner. It is also possible to discriminate between small mutations and large deletions by checking the presence of the linked marker, neo. As a model experiment, we prepared an uncloned pool of rat fibroblast cells (CREF) infected with this virus and irradiated them with increasing doses of ultraviolet light. Dose-dependent increases in the number of ACV-resistant colonies were observed. Structural analysis of the HSV-TK gene in these clones revealed point mutations or small deletions in the majority of the cases. Since it requires no pre-existing genetic markers in the host cells, this system may be used for a wide variety of mammalian cells and provides a useful tool to assess both their susceptibility to various mutagens and their genomic instability
- リンク情報
- ID情報
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- DOI : 10.1016/0165-1161(95)90075-6
- ISSN : 0165-1161
- J-Global ID : 200902164389051076
- Web of Science ID : WOS:A1995QW61200012