Phenotypic conversion of TK-deficient cells following electroporation of functional TK enzyme.
The ability to phenotypically rescue a mutant (Rat-3, thymidine kinase-deficient) cell line by electroporation of functional TK enzyme has been investigated. Extracts of electroporated cells showed a 35-fold increase in TK enzyme levels under conditions where greater than 90% of the cells remained viable. The electroporated enzyme was intracellular, as demonstrated by the fact that cells were able to utilize exogenous [3H]thymidine for DNA synthesis. By in situ autoradiography, 82% of electroporated cells contained functional enzyme and incorporated [3H]thymidine into DNA. Thus, this technique can efficiently provide a missing metabolic function to cultured mammalian cells.
Dagher
S F
SF
Graduate Program in Genetics, Michigan State University, East Lansing 48824-1101.
Conrad
S E
SE
Werner
E A
EA
Patterson
R J
RJ
D000818:Animals; D001345:Autoradiography; D002460:Cell Line; D003584:Cytological Techniques; D004558:Electric Stimulation; D006801:Humans; D007700:Kinetics; D008856:Microscopy, Fluorescence; D010641:Phenotype; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013936:Thymidine / Q000378:metabolism; D013937:Thymidine Kinase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D013937:Thymidine Kinase; D013936:Thymidine