微生物DNA与培养的哺乳动物细胞的相互作用。供体DNA与细胞表面的结合。

Interaction of microbial DNA with cultured mammalian cells. Binding of the donor DNA to the cell surface.

作者信息

Ehrlich M, Sarafyan L P, Myers D J

出版信息

Biochim Biophys Acta. 1976 Dec 13;454(3):397-409. doi: 10.1016/0005-2787(76)90266-5.

Abstract

Cultured fibroblasts grown in monolayer were incubated for a short time with radioactively labeled microbial DNA and diethylaminoethyl-dextran (DEAE-dextran), poly-L-lysine, or calcium phosphate, agents previously demonstrated by others to markedly enhance transfection. Immediately after such treatment of 1 by 10(-6)-1.5 by 10(6) cells with DNA, approx. 0.05-0.15 mug of donor DNA representing 10-30% of the input DNA became cell associated. In contrast, when the cells are similarly treated with only a DNA solution approx. 0.5-5% of the donor DNA was retained by the cells. More than 95% of the cell-associated donor DNA was shown to be bound to the surface of cells treated with polycation. It was also shown that in the absence of polycation treatment, most of the cell-associated donor DNA was bound to the cell surface.

摘要

将单层培养的成纤维细胞与放射性标记的微生物DNA和二乙氨基乙基葡聚糖（DEAE-葡聚糖）、聚-L-赖氨酸或磷酸钙一起孵育较短时间，其他人先前已证明这些试剂能显著增强转染效果。在用DNA对1×10⁻⁶至1.5×10⁶个细胞逐个进行此类处理后，立即有大约0.05 - 0.15微克的供体DNA（占输入DNA的10 - 30%）与细胞结合。相比之下，当细胞仅用DNA溶液进行类似处理时，细胞仅保留了大约0.5 - 5%的供体DNA。结果表明，超过95%与细胞结合的供体DNA与用聚阳离子处理的细胞表面结合。还表明，在没有聚阳离子处理的情况下，大多数与细胞结合的供体DNA也与细胞表面结合。