摘要
本文用改良的 Giuffrida法分离纯化大鼠大脑皮层细胞核,产率为41—52％,具有很高的体外转录活性。参照Blatti硫酸铵浓度法建立了大鼠大脑皮层细胞核体外转录模型,能分别测定真核基因三类 RNA聚合酶转录活性,并对有关问题进行了讨论,指出本系统具有操作简便、省时、省实验材料等优点,适用于大脑皮层细胞核体外转录调控的研究。
This paper deals with establishing a transcriptional model in which the transcriptional activities of RNA Pol Ⅰ, Ⅱ, Ⅲ in intact nucleus could be determined separately in vitro. With modified Giuffrida method, the nuclei isolated from rat cerebral cortex were purified in good yields ranging from 41 to 52％ and high purity judging with phase-contrast microscopy. According to the method described by Blatti, RNA synthsis were studied under high or low (240 or 50 mmol/L (NH)_2SO_4) ionic strength condition. The inhibited and residual transcription in high-salt system and presence of α-amanitin represented the activities engaged by RNA Pol Ⅱ and Pol Ⅲ independently, the transcription in low-salt system containing α-amanitin was mainly responsible for RNA PolⅠactivities. In addition, some factors effecting on the nuclei transcription in vitro were also evaluated. It was concluded that the reported model had advantages being easy to operated, saving materials, and approching cell physiological conditions, it was suitable for studying transcriptional mechanism of cerebral cortex neurons.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1990年第1期45-49,共5页
Progress In Biochemistry and Biophysics
关键词
RNA聚合酶
转录
大脑皮层
细胞核
RNA polymerase (RNA Pol), transcription, cerebral cortex, nuclei
