Transfer of herpes simplex virus thymidine kinase synthesized in bacteria by a high-expression plasmid to tissue culture cells by protoplast fusion.
The introduction of a protein into living tissue culture cells may permit the in vivo study of functions of the protein. We have previously described a high-efficiency-expression plasmid, pHETK2, containing the herpes simplex virus type 1 thymidine kinase (TK) gene which, upon temperature induction, causes TK to be synthesized as greater than 4% of the bacterial protein. In this report we show that enzymatically active TK was transferred to mouse Ltk- cells by polyethylene glycol-mediated fusion with protoplasts prepared from bacteria containing induced levels of TK. The presence of TK in the Ltk- cells was detected by the incorporation of [3H]thymidine into cell nuclei as measured by autoradiography.
D000818:Animals; D001345:Autoradiography; D002459:Cell Fusion; D002467:Cell Nucleus / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D007739:L Cells / Q000201:enzymology; D051379:Mice; D010957:Plasmids; D011523:Protoplasts / Q000201:enzymology; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics*; D014316:Tritium
D014316:Tritium; D013937:Thymidine Kinase