Brown D B, Hanks S K, Murphy E C, Rao P N
Exp Cell Res. 1985 Jan;156(1):251-9. doi: 10.1016/0014-4827(85)90279-4.
A modification of polyethylene glycol-mediated cell fusion procedure has been described and standardized for red blood cell mediated microinjection of proteins into cells in suspension. Using this procedure, proteins are routinely introduced into 50% of the target cells. We have applied this microinjection procedure to introduce cytoplasmic extracts obtained from S-phase HeLa cells into HeLa cells synchronized in G1. This experiment resulted in an accelerated entry of the G1 cells into S phase as measured by the incorporation of [3H]thymidine. This technique may provide a means to identify and characterize the S-phase factors responsible for the induction of DNA synthesis.
已经描述并标准化了一种聚乙二醇介导的细胞融合程序的改良方法,用于红细胞介导的将蛋白质显微注射到悬浮细胞中。使用该程序,蛋白质通常被导入50%的靶细胞。我们已应用这种显微注射程序,将从S期HeLa细胞获得的细胞质提取物导入同步于G1期的HeLa细胞。通过[3H]胸苷掺入量测定,该实验导致G1期细胞加速进入S期。这项技术可能提供一种手段,来鉴定和表征负责诱导DNA合成的S期因子。