Efficient expression of foreign genes in CHO DHFR(-) cells by electroporation.
DHFR-deficient Chinese hamster ovary (CHO DHFR(-)) cells are the most popular mammalian expression system for inducible amplification of transgene. In order to obtain more stable transfected CHO DHFR(-) cell clones, transfection efficiency of electroporation under different conditions were systemically investigated using plasmid pSV-beta-Gal as reporter gene. Transfection efficiency was proportionally increased with pulse duration and number of pulse applied. In addition, higher transfection efficiency was found in high salt extracellular solution (Berg's and Hank's buffers) than in intracellular solution (cytomix buffer) under the same electroporation condition. The highest transfection efficiency in examined conditions was about 1 in 350 cells (or 0.289%) when cells were electroporated with twice pulses at 400V, 375microF. The present study offers an optimized guideline for introducing exogenous DNA into CHO DHFR(-) cells by electroporation.
Lin
Wen-Zhen
WZ
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China. linwenzhen@msn.com
Lee
Susanna S T
SS
Cheung
Wing-Tai
WT
D000818:Animals; D002021:Buffers; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004526:Efficiency; D018274:Electroporation / Q000379:methods; D015870:Gene Expression; D005796:Genes; D017930:Genes, Reporter; D005822:Genetic Vectors; D013762:Tetrahydrofolate Dehydrogenase / Q000172:deficiency / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D002021:Buffers; D013762:Tetrahydrofolate Dehydrogenase