Transcription of viral genes by RNA polymerase II in nuclei isolated from adenovirus 2 transformed cells.
Transcription of viral genes by RNA polymerase II in nuclei isolated from adenovirus 2 transformed cells.
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RNA 聚合酶 II 在从腺病毒 2 转化细胞中分离的细胞核中转录病毒基因。
DOI：
10.1021/bi00604a028
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发表时间：
1978
期刊：
影响因子：
2.9
通讯作者：
R. Roeder
中科院分区：
文献类型：
--
作者：
G. Bitter;R. Roeder
Grant A. Bitter and Robert G. Roeder* abstract: An in vitro transcription system utilizing nuclei isolated from adenovirus type 2 transformed cells has been developed. Cloned cell lines which contain viralDNA se-quences complementary to only the left 14% of the adenovirus genome [Gallimore, P. H., Sharp, P. A., and Sambrook, J.(1974), J. Mol. Biol. 89, 49] have been used in the present study. The sensitivity to-amanitin of RNA synthesis in iso-lated nuclei from B1 cells demonstrates the presence of the three classes of RNA polymerase which are characteristic of eukaryotic cells. In the presence of Mn2+, the endogenous RNA polymerase activity (predominantly RNA polymerase II) shows a biphasic ammonium sulfate activation profile. At the lower salt optimum (0.03 M ammonium sulfate), synthesis continues for 40 min at 30 C. Pulse-chase experiments demonstrate that the in vitro product is stable for an extended period of time after synthesis. In addition, studies with heparin indicate that negligible initiation of RNA chains occurs in the isolated nuclei. Thus, the observed synthesis appears to consist entirely of elongation of RNA chains which were initiated in
