The dynamics of gene expression in transiently transferred mammalian cells were analyzed using the green fluorescent protein (Gfp), along with flow cytometric analysis. The highest fraction of transfectants was measured as lipid-DNA complexes form with a certain amount of lipid and DNA for chinese hamster ovary cells and NIH/3T3 cells. Results indicate that green fluorescence is a quantitative measure of intracellular Gfp in single cells in spite of the dynamics of post-transitional modifications involved in the conversion of expressed protein into its fluorescence form. A structured model has been developed to describe the observed kinetics of gene expression and fluorophore formation.
Bibliographical noteFunding Information:
We thank R. Tsien for kindly providing the Gfp-encoding genes. We also thank D. Bernlohr and C. Whitley for providing us with the cell lines used in this study. We are grateful to S. Fahrenkrug and P.B. Hackett for their helpful advice. This work has been supported in part by the National Science Foundation (BES-941438.5). We are grateful to D. Jackson for helping in the preparation of this manuscript.
- CHO cell culture
- Flow cytometry
- Green fluorescent protein
- Structured model
- Transient gene expression