Cytotoxic effect of Drosophila deoxynucleoside kinase gene on replicating plasmid in HeLa cells.
To enhance the levels of transgene expression from plasmid-based nonviral vectors, replicating plasmids containing the SV40 origin and the SV40 large T antigen gene, as a model replicating unit, were constructed. The replicating luciferase plasmid DNA produced the luciferase protein more efficiently than the non-replicating luciferase plasmid DNA, as expected. Surprisingly, the introduction of the replicating plasmid DNA containing the Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) gene was highly cytotoxic and caused cell death without nucleoside analogs. Our results confirm that transgenes on a replicating plasmid represent an excellent tool for effective protein production and suggest that efficient production of the Dm-dNK protein in tumor cells could be an attractive cancer therapy.
Ito
Mana
M
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Suda
Yusuke
Y
Harashima
Hideyoshi
H
Kamiya
Hiroyuki
H
D000818:Animals; D001483:Base Sequence; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D017931:DNA Primers; D004331:Drosophila melanogaster / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D010957:Plasmids; D019076:Transgenes
D017931:DNA Primers; D008156:Luciferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C020018:deoxyribonucleoside kinases; D053148:Caspase 3; D053179:Caspase 7