Nicolau C, Sene C
Biochim Biophys Acta. 1982 Oct 11;721(2):185-90. doi: 10.1016/0167-4889(82)90067-2.
The pBR322 plasmid containing the sequence encoding beta-lactamase, the enzyme conferring resistance to ampicillin, was encapsulated in liposomes of different phospholipid composition and incubated with synchronized cells. In mitotic cells as compared to cells synchronized in G1, twice as many exogeneous DNA molecules were found associated with the cell nuclear DNA, when fluid, neutral liposomes were used. These liposomes are taken up by the cells mainly via endocytosis. When fluid, negatively charged liposomes were used as carriers about the same number of exogenous DNA molecules were found associated with the nuclear DNA both in mitotic and in G1-synchronized cells. The efficiency for gene transfer of liposomes entering the cells by different mechanisms was further studied and expressed both by the fraction of the radioactive plasmid associated with the nuclear DNA and by the level of the beta-lactamase activity detected in the transfected cells. It appears that liposomes entering the cells mainly via an energy-dependent mechanisms are more efficient for this type of DNA transfer.
携带编码β-内酰胺酶(一种赋予对氨苄青霉素抗性的酶)序列的pBR322质粒被包裹在不同磷脂组成的脂质体中,并与同步化细胞一起孵育。当使用流动性中性脂质体时,与G1期同步化细胞相比,在有丝分裂细胞中发现与细胞核DNA相关的外源DNA分子数量是其两倍。这些脂质体主要通过内吞作用被细胞摄取。当使用流动性带负电荷的脂质体作为载体时,在有丝分裂细胞和G1期同步化细胞中,发现与核DNA相关的外源DNA分子数量大致相同。通过不同机制进入细胞的脂质体的基因转移效率得到了进一步研究,并通过与核DNA相关的放射性质粒的比例以及在转染细胞中检测到的β-内酰胺酶活性水平来表示。似乎主要通过能量依赖机制进入细胞的脂质体对于这种类型的DNA转移更有效。