DNA-mediated gene transfer of a circular plasmid into murine cells.

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Abstract
DNA-mediated gene transfer was used to introduce a recombinant plasmid (pBR322) containing the human .beta.-globin gene (H.beta. 1) into cells of a mouse tissue culture line, Ltk- (thymidine kinase deficient neoplastic fibroblast cell). 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 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 MW DNA. DNA from 3 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.