Iino T, Furusawa M, Furusawa I, Obinata M
Exp Cell Res. 1983 Oct 15;148(2):475-80. doi: 10.1016/0014-4827(83)90168-4.
Fusion of red cell ghosts containing foreign materials with cells results in the introduction of the materials into the cells (red cell-mediated microinjection). Until now, 'two-step dialysis' has mainly been used for trapping proteins in the ghosts. Large-sized materials such as DNA, however, are rarely trapped in the ghosts, since the holes in the red cell membrane caused by osmotic shock are too small for such materials to pass through. In this study, we improved the trapping technique. Some of the Hind III fragments of lambda phage DNA as well as proteins could be trapped in the ghosts when the mixture of these materials and red cells were frozen at -80 degrees C for a short period followed by quick thawing. Red cell-mediated microinjection using ghosts containing plasmid pBR322 linked with a Herpes simplex viral thymidine kinase (tk) gene brought about transformation of tk-defective L cells, the efficiency of transformation was 1 out of 20 000-60 000 cells fused with the ghosts.
含有异物的红细胞血影与细胞融合会将这些物质引入细胞（红细胞介导的显微注射）。到目前为止，“两步透析”主要用于将蛋白质捕获到血影中。然而，诸如DNA等大分子物质很少能被捕获到血影中，因为渗透休克导致的红细胞膜上的孔对于这类物质来说太小，无法穿过。在本研究中，我们改进了捕获技术。当这些物质与红细胞的混合物在-80℃短暂冷冻后快速解冻时，λ噬菌体DNA的一些Hind III片段以及蛋白质能够被捕获到血影中。使用含有与单纯疱疹病毒胸苷激酶（tk）基因相连的质粒pBR322的血影进行红细胞介导的显微注射，导致tk缺陷的L细胞发生转化，转化效率为与血影融合的20000 - 60000个细胞中有1个发生转化。
