Fugene 6——a new approach to induce the gene transfection in high efficiency for eukaryotic cells in vitro / 免疫学杂志

ABSTRACT

Objective　To develop a new method to induce the gene transfection in high efficiency for eukaryotic cells in vitro. Methods　 Four kinds of p14ARF gene primarily deleted human carcinoma cell line including H460,A549,U251,and PC-3 were transfected with the human p14ARF expression vector (pCI-neo-p14ARF) by using the new nonliposomal transfection reagent Fugene 6. The efficiency of gene transfer was determined by screening the cells in G418. Results　 After 21 days' selection, G418-resistant clones were shown in all the transfected plate. PCR product of p14ARF gene was positive in all the G418-resistant clones. Cytotoxicity of Fugene 6 was detected. The cell proliferation activity was not affected when it was cultured in a high dose of Fugene 6. Conclusion　 These results demonstrate that Fugene 6 is a rapid, feasible, reproducible, and noncytotoxic gene transfection approach for eukaryotic expression vector in vitro.