Extrachromosomal unequal homologous recombination and gene conversion in simian kidney cells: effects of UV damage |
| |
| Affiliation: | 1. Michael G. DeGroote School of Medicine, Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON L8N 3Z5, Canada;2. Department of Medicine, Division of Hematology, Schulich School of Medicine, University of Western Ontario, London, Ontario N6A 3K7, Canada;3. Hematology Exploration and Applications in Leukemia (HEAL) Program, Hamilton, ON, Canada;4. Department of Oncology, McMaster University, Hamilton, ON L8S 4L8, Canada;5. Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4L8, Canada;1. State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Normal University, Changsha, 410081, China;2. State Key Laboratory of Developmental Biology of Freshwater Fish, Engineering Research Center of Polyploid Fish Reproduction and Breeding of the State Education Ministry, Changsha, 410081, China;3. College of Life Sciences, Hunan Normal University, Changsha, 410081, China;4. Yueyang Tieshan Water Supply Project Affairs Center, Yueyang, 41699, China;5. Yueyang Tieshan Water Supply Project Affairs Center, Tieshan Reservoir Management Office, Yueyang, 414114, China;1. Dipartimento di Scienze della Terra e del Mare (DiSTeM), Università di Palermo, Via Archirafi 20, I-90123 Palermo, Italy;2. CoNISMa, Piazzale Flaminio 9, 00197 Roma, Italy;3. Université Côte d’Azur, CNRS, FRE 3729 ECOMERS, Parc Valrose 28, Avenue Valrose, 06108 Nice, France;4. Institut de Ciències del Mar, CSIC, Psg. Marítim de la Barceloneta 37-49, 08003 Barcelona, Spain;1. Chromatin and Disease Group, Cancer Epigenetics and Biology Programme (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L''Hospitalet de Llobregat, Barcelona, Spain;2. Autoinflammatory Diseases Clinical Unit and Vasculitis Research Unit, Department of Autoimmune Diseases, Hospital Clinic, IDIBAPS, Barcelona, Spain;3. Internal Medicine, Autoimmune and Systemic Diseases Services, Hospital Vall d''Hebron, Barcelona, Spain;4. Allergy Department, Hospital Universitari Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona, Spain;5. Department of Pediatrics, Hospital Universitari Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona, Spain;6. Department of Immunology-CDB, Hospital Clinic, IDIBAPS, Barcelona, Spain |
| |
| Abstract: | Shuttle plasmid vectors containing the SV40 origin of replication and tandem neo genes with distally placed non-overlapping deletions were used to study the effects of DNA damage on extrachromosomal homologous recombination in simian kidney cells. DNA was introduced into COS7 cells by a lipofectin-mediated transfection procedure and recombination was assessed by analyzing the structure of plasmids. Recombinational events observed included unequal homologous recombination (triplication), gene conversion, double reciprocal recombination, deletion (pop-outs), gene amplification (4–6 copies), and multimerization. Triplication, an event that previously had not been reported in association with extrachromosomal recombination, predominated in experiments with undamaged vectors. The recombination frequency (NeoR/AmpR) of vectors randomly damaged by UV irradiation was essentially unchanged; however, the relative number of triplication events decreased significantly. Selective damage in one of the two neo genes increased the relative frequency of gene conversion. The experimental system developed for use in this study detects all major homologous recombination events observed in chromosomal direct repeat sequences in mammalian cells and yeast and should prove valuable for future studies of homologous recombination in mammalian cells. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
