Mory Y Y, Gefter M L
Nucleic Acids Res. 1977 Jun;4(6):1739-57. doi: 10.1093/nar/4.6.1739.
Nuclei isolated from mouse myeloma cells grown in tissue culture are capable of synthesizing RNA for prolonged periods of time. Addition of cytoplasmic extracts to the system stimulates slightly the rate and prolongs the time of synthesis. As judges by sedimentation in SDS and in formamide gradients, the size of the RNA synthesized is heterogeneous from smaller than 10S to larger than 45S, thus resembling in vivo made RNA. The characteristics of some of the RNA are in keeping with those expected to be for mRNA. Fifty percent of the RNA synthesis is sensitive to alpha-amanitin. After an incubation of two hours in the absence of alpha-amanitin about 10 percent of the newly synthesized RNA is found outside of the nuclei; it sediments with a broad distribution at 18S. A considerable fraction of the RNA that is released from nuclei in vitro can promote the formation of polyribosomes, and contains molecules that are polyadenylated and "capped".
从组织培养中生长的小鼠骨髓瘤细胞分离出的细胞核能够长时间合成RNA。向该系统中添加细胞质提取物会略微刺激合成速率并延长合成时间。通过在SDS和甲酰胺梯度中的沉降判断,合成的RNA大小从小于10S到大于45S不等,因此类似于体内合成的RNA。某些RNA的特征与预期的mRNA特征相符。50%的RNA合成对α-鹅膏蕈碱敏感。在没有α-鹅膏蕈碱的情况下孵育两小时后,约10%新合成的RNA在细胞核外被发现;它在18S处有广泛分布的沉降。体外从细胞核释放的相当一部分RNA能够促进多核糖体的形成,并且包含多聚腺苷酸化和“加帽”的分子。