外源RNA聚合酶在染色体细胞学制片上进行RNA合成。
RNA synthesis by exogenous RNA polymerase on cytological preparations of chromosomes.
作者信息
Sederoff R, Clynes R, Poncz M, Hachtel S
出版信息
J Cell Biol. 1973 May;57(2):538-50. doi: 10.1083/jcb.57.2.538.
Abstract
Cytological preparations of Drosophila polytene chromosomes serve as templates for RNA synthesis carried out by exogenous RNA polymerase (Escherichia coli). Incorporation of labeled ribonucleoside triphosphates into RNA may be observed directly by autoradiography. Because of the effects of rifampicin, actinomycin D, ribonuclease, high salt, and the requirement for all four nucleoside triphosphates, we conclude that the labeling observed over chromosomes is due to DNA-dependent RNA polymerase activity. Using this method, one can observe RNA synthesis in vitro on specific chromosome regions due to the activity of exogenous RNA polymerase. We find that much of the RNA synthesis in this system occurs on DNA sequences which appear to be in a nondenatured state.
摘要
果蝇多线染色体的细胞学制剂可作为外源RNA聚合酶（大肠杆菌）进行RNA合成的模板。通过放射自显影可直接观察到标记的核糖核苷三磷酸掺入RNA的情况。由于利福平、放线菌素D、核糖核酸酶、高盐的影响以及对所有四种核苷三磷酸的需求，我们得出结论，在染色体上观察到的标记是由于依赖DNA的RNA聚合酶活性所致。使用这种方法，由于外源RNA聚合酶的活性，可以在体外观察特定染色体区域的RNA合成。我们发现该系统中的许多RNA合成发生在似乎处于非变性状态的DNA序列上。
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