Lévesque J P, Sansilvestri P, Hatzfeld A, Hatzfeld J
Laboratoire C.N.R.S., I.C.I.G. Hôpital Paul-Brousse, Villejuif, France.
Biotechniques. 1991 Sep;11(3):313-4, 316-8.
We describe a defined medium that allows efficient DNA transfections in COS cells and transient expression of the corresponding recombinant protein in serum-free conditions. With a modified DEAE-dextran/chloroquine method, we obtained 80% more transfected cells expressing the recombinant human interleukin-2 receptor than with transfection with cationic liposomes, one of the most efficient techniques to date. The absence of serum in the culture medium should reduce subsequent purification steps for production of recombinant mammalian proteins. Moreover, it should allow investigations dealing with the role of serum or other exogenous factors on mRNA stability or post-translation events during protein synthesis.
我们描述了一种特定培养基,其能在无血清条件下实现COS细胞中的高效DNA转染以及相应重组蛋白的瞬时表达。通过改良的DEAE-葡聚糖/氯喹方法,相较于目前最有效的技术之一——阳离子脂质体转染,我们获得了多80%表达重组人白细胞介素-2受体的转染细胞。培养基中无血清应能减少重组哺乳动物蛋白生产后续的纯化步骤。此外,这应有助于开展关于血清或其他外源因子对mRNA稳定性或蛋白质合成过程中翻译后事件作用的研究。