Givol I, Tsarfaty I, Resau J, Rulong S, da Silva P P, Nasioulas G, DuHadaway J, Hughes S H, Ewert D L
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201.
Cell Growth Differ. 1994 Apr;5(4):419-29.
A complementary DNA for human bcl-2 was cloned into the replication competent avian retrovirus vector RCASBP, and the resulting virus was used to express human Bcl-2 protein at high levels in chicken embryo fibroblasts. The expression of Bcl-2 did not transform or significantly alter the longevity of the chicken embryo fibroblasts in the presence of normal amounts of serum. However, the expression of Bcl-2 blocked c-Myc-induced apoptosis in these cells. Fractionation of the infected chicken embryo fibroblasts indicated that the protein was distributed equally between nuclear and high density cytoplasmic membranes. Immunofluorescence analysis by confocal microscopy and immunoelectron microscopy showed that the Bcl-2 protein was primarily associated with the nuclear membrane and with the endoplasmic reticulum. Reduced amounts of the protein were associated with other membranes in the cytoplasm. These data show that, in this system, the Bcl-2 protein associates with the nuclear membrane and intracytoplasmic membranes but is not preferentially associated with mitochondria.
将人类bcl - 2的互补DNA克隆到具有复制能力的禽逆转录病毒载体RCASBP中，然后用所得病毒在鸡胚成纤维细胞中高水平表达人类Bcl - 2蛋白。在正常量血清存在的情况下，Bcl - 2的表达并未使鸡胚成纤维细胞发生转化或显著改变其寿命。然而，Bcl - 2的表达阻断了这些细胞中c - Myc诱导的凋亡。对感染的鸡胚成纤维细胞进行分级分离表明，该蛋白在核膜和高密度细胞质膜之间均匀分布。通过共聚焦显微镜和免疫电子显微镜进行的免疫荧光分析表明，Bcl - 2蛋白主要与核膜和内质网相关。该蛋白在细胞质中与其他膜相关的量较少。这些数据表明，在该系统中，Bcl - 2蛋白与核膜和胞内膜相关，但并非优先与线粒体相关。
