Kinetic analysis of protein production after DNA transfection.
The production of an exogenous protein by the transfection of a plasmid DNA encoding the protein was kinetically analyzed, to determine the efficiency of the transfection. Cultured NIH3T3 or HeLa cells, and the luciferase protein were used as a model system in this experiment. The findings indicate that at least a 8x10(4)- and 4x10(3)-fold molar amounts of luciferase protein was produced from one copy of the plasmid DNA molecule in NIH3T3 and HeLa cells, respectively. The rate of elimination of luciferase activity upon DNA transfection was smaller than that for the luciferase protein itself (k(el) for DNA transfection<k(el) for the luciferase protein), suggesting that a decrease in intranuclear active DNA was the main determinant of the elimination rate in this case. A preliminary pharmacokinetic model is proposed, based on the results obtained.
Yamada
Yuma
Y
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Kamiya
Hiroyuki
H
Harashima
Hideyoshi
H
D000818:Animals; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008954:Models, Biological; D041681:NIH 3T3 Cells; D010957:Plasmids; D014176:Protein Biosynthesis / Q000235:genetics*; D014162:Transfection
D004247:DNA; D008156:Luciferases