Tatsuka M, Orita S, Yagi T, Kakunaga T
Department of Oncogene Research, Osaka University, Japan.
Exp Cell Res. 1988 Sep;178(1):154-62. doi: 10.1016/0014-4827(88)90386-2.
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.
我们开发了一种改良的、可重复且高效的方法，通过施加电场然后用丁酸钠处理，将克隆基因导入哺乳动物细胞。由抗生素（G418）抗性基因和猴病毒40（SV40）早期启动子组成的质粒pSV2-neo，通过电穿孔法的转染频率高于磷酸钙DNA沉淀法。电穿孔后用丁酸钠处理显著提高了包括人皮肤成纤维细胞在内的各种细胞系和原代培养细胞的转染频率。当通过电穿孔将氯霉素乙酰转移酶（乙酰辅酶A；氯霉素O3-乙酰转移酶，CAT，EC 2.3.1.28）基因导入BALB/c 3T3细胞时，用丁酸钠处理也增加了该基因的瞬时表达。电穿孔结合丁酸钠处理是一种用于在培养的哺乳动物细胞中稳定和瞬时进行外源基因生化转化的改进方法。
