DNA-mediated gene transfer of a circular plasmid into murine cells.
We have used DNA-mediated gene transfer to introduce a recombinant plasmid containing the human beta-globin gene (H beta 1) into cells of a mouse tissue culture line, Ltk-. DNA isolated from independent transfer lines was analyzed by restriction endonuclease digestion, gel electrophoresis, modified Southern blotting, and filter hybridization using H beta 1 as a probe. H beta 1 sequences were present in 80% of the lines at 1-30 copies per cell. Many of the lines gave a hybridization pattern indicative of H beta 1 sequences integrated into high molecular weight DNA. DNA from three cell lines, digested with several restriction enzymes, produced a pattern providing evidence for the presence of circular H beta 1 molecules in the murine recipient cells.
D000818:Animals; D002460:Cell Line; D004261:DNA Replication; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004270:DNA, Circular / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant; D005111:Extrachromosomal Inheritance; D005796:Genes; D005914:Globins / Q000235:genetics; D051379:Mice; D010957:Plasmids; D013937:Thymidine Kinase / Q000235:genetics; D014170:Transformation, Genetic
D004270:DNA, Circular; D004274:DNA, Recombinant; D005914:Globins; D013937:Thymidine Kinase; D004262:DNA Restriction Enzymes