Convenient plasmid for extrachromosomal DNA recombination in mouse cells.
We constructed a convenient plasmid for DNA recombination assay. The plasmid, pMR1, contains a double prokaryotic terminator to decrease the background and two unique restriction enzyme sites on both sides of the double terminator to allow for easy construction. The assay is capable of selecting the bacterial cells containing recombined plasmid DNA on a selective plate containing ampicillin and chloramphenicol. We adapted pMR1 for V(D)J recombination and homologous recombination and detected both types of recombination in murine PreB cell line. As pMR1 has the double terminator, background on the selective plate decreases effectively and we select only the recombined clones. We consider the vector, pMR1, to be convenient for the analysis of homologous and non-homologous recombinations.
Kameyama
K
K
Department of Chemistry II, Faculty of Science, Hokkaido University, Sapporo, Japan.
Mochizuki
M
M
Tanaka
K
K
Sugimoto
K
K
D000011:Abelson murine leukemia virus / Q000235:genetics; D000667:Ampicillin / Q000494:pharmacology; D000668:Ampicillin Resistance / Q000235:genetics; D000818:Animals; D001402:B-Lymphocytes; D001483:Base Sequence; D002461:Cell Line, Transformed; D002701:Chloramphenicol / Q000494:pharmacology; D002702:Chloramphenicol Resistance / Q000235:genetics; D004247:DNA / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013728:Terminator Regions, Genetic; D014162:Transfection; D014170:Transformation, Genetic / Q000187:drug effects
D007143:Immunoglobulin Heavy Chains; D009838:Oligodeoxyribonucleotides; D021382:Protein Sorting Signals; D002701:Chloramphenicol; D000667:Ampicillin; D004247:DNA