DNA transformation of a pluripotent mouse embryonal stem cell line with a dominant selective marker.
The mouse embryonal stem cell line BLC 1 growing on feeder layer was treated with a calcium phosphate/DNA precipitate prepared with DNA of plasmid pAG60 which harbors the Tn5-derived neo gene, thus encoding resistance to G418, an aminoglycoside antibiotic. Transfection performed on feeder layer resulted in the formation of G418-resistant clones T1 and T2/K26. The stable integration of the transformed neo gene was confirmed by dot hybridization in all descendant cultures of clones T1 and T2/K26 as well as in the tumors derived from them. In vivo and in vitro differentiation revealed the pluripotent status of the transformants. Tumors derived from T1 and T2/K26 contained various tissues with derivatives of all three primary germ layers.
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002454:Cell Differentiation; D002469:Cell Separation; D002999:Clone Cells; D004247:DNA / Q000494:pharmacology*; D004351:Drug Resistance; D005799:Genes, Dominant; D005819:Genetic Markers; D005839:Gentamicins / Q000494:pharmacology; D006824:Hybridization, Genetic; D051379:Mice; D013234:Stem Cells / Q000166:cytology*; D013724:Teratoma / Q000503:physiopathology; D014170:Transformation, Genetic
D005819:Genetic Markers; D005839:Gentamicins; D004247:DNA; C010680:antibiotic G 418