Direct transfection of viral and plasmid DNA into the liver or spleen of mice.
A method for the direct transfection of polyoma viral DNA and polyoma-plasmid recombinant DNA into the liver or spleen of newborn or adult mice was developed. Calcium phosphate-precipitated DNA was injected directly into mouse organs in combination with hyaluronidase and collagenase. Transfected DNA was shown to replicate at moderate efficiency, relative to direct infection of organs with virus. Transfection with viral DNA rapidly led to an acute infection. A polyoma-bacterial plasmid recombinant DNA also was shown to replicate when transfected into mice. With this plasmid, however, genomic-length polyoma DNA rapidly recombined away from the bacterial component and replicated as viral DNA. This method should allow the direct determination of the biological activity of a cloned DNA within a mouse organ.
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics*; D008099:Liver / Q000378:metabolism*; D051379:Mice; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011120:Polyomavirus / Q000235:genetics*; D013154:Spleen / Q000378:metabolism*; D014162:Transfection
D004279:DNA, Viral; D004262:DNA Restriction Enzymes