Weidner D A, Winicov I
Cell Biophys. 1983 Dec;5(4):255-64. doi: 10.1007/BF02788624.
We have been able to obtain significant levels of apparently in vitro-initiated beta-globin transcripts from isolated nuclei using mouse Friend cells. The level of hybrid selected beta-globin transcripts was increased 3-9-fold in nuclei after the cells were induced with dimethylsulfoxide. The newly initiated transcripts were detected through incorporation of gamma-thio ATP and isolation by Hg-agarose chromatography. The 5'-terminus was identified by recovery of the thio tetraphosphate in hydrolysis products from in vitro initiated RNA using gamma-35S ATP.
我们已经能够使用小鼠Friend细胞从分离的细胞核中获得显著水平的明显由体外起始的β-珠蛋白转录本。在用二甲基亚砜诱导细胞后,细胞核中杂交选择的β-珠蛋白转录本水平增加了3至9倍。通过掺入γ-硫代ATP并经汞琼脂糖层析分离来检测新起始的转录本。使用γ-35S ATP通过回收体外起始RNA水解产物中的硫代四磷酸来鉴定5'-末端。