An improved method of electroporation for introducing biologically active foreign genes into cultured mammalian cells.
We have developed a modified, reproducible, and efficient method for introducing cloned genes into mammalian cells by using an electric field followed by treatment with sodium butyrate. Transfection frequencies with plasmid pSV2-neo, consisting of an antibiotic (G418) resistance gene and simian virus 40 (SV40) early promoter, by electroporation were higher than those by calcium phosphate DNA precipitation. Treatment with sodium butyrate following electroporation significantly increased the frequency of transfection in various types of cell lines and primary cultured cells including human skin fibroblasts. Treatment with sodium butyrate also increased the transient expression of the gene for chloramphenicol acetyltransferase (acetyl-CoA; chloramphenicol O3-acetyltransferase, CAT, EC 2.3.1.28) when the gene was introduced into BALB/c 3T3 cells by electroporation. Electroporation combined with sodium butyrate treatment is an improved method for stable and transient biochemical transformation of foreign genes in cultured mammalian cells.
Tatsuka
M
M
Department of Oncogene Research, Osaka University, Japan.
Orita
S
S
Yagi
T
T
Kakunaga
T
T
D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002130:Calcium Phosphates / Q000494:pharmacology; D002452:Cell Count; D002460:Cell Line; D002463:Cell Membrane Permeability; D004247:DNA / Q000187:drug effects / Q000502:physiology; D004558:Electric Stimulation; D005990:Glycerol / Q000494:pharmacology; D006801:Humans; D008322:Mammals; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012997:Solvents; D013696:Temperature; D014162:Transfection
D002087:Butyrates; D002130:Calcium Phosphates; D011993:Recombinant Fusion Proteins; D012997:Solvents; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D020148:Butyric Acid; C485838:calcium phosphate, monobasic, anhydrous; D004247:DNA; C020243:calcium phosphate; C485829:calcium phosphate, dibasic, anhydrous; D005990:Glycerol