Efficient expression of foreign genes in CHO DHFR- cells by electroporation
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AbstractDHFR-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 I in 350 cells (or 0.289%) when cells were electroporated with twice pulses at 400 V, 375 mu F. The present study offers an optimized guideline for introducing exogenous DNA into CHO DHFR- cells by electroporation. (C) 2009 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.
All Author(s) ListLin WZ, Lee SST, Cheung WT
Journal nameBiologicals
Detailed descriptionTo ORKTS: doi: 10.1016/j.biologicals.2009.03.003
Volume Number37
Issue Number5
Pages277 - 281
LanguagesEnglish-United Kingdom
KeywordsCHO DHFR- cells; Electroporation; Transfection
Web of Science Subject CategoriesBiochemical Research Methods; BIOCHEMICAL RESEARCH METHODS; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; BIOTECHNOLOGY & APPLIED MICROBIOLOGY; Pharmacology & Pharmacy; PHARMACOLOGY & PHARMACY

Last updated on 2020-28-09 at 01:14