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利用噬菌体T4蛋白进行DNA复制。通过互补分析对T4基因41、45、44和62编码的蛋白进行纯化。

DNA replication with bacteriophage T4 proteins. Purification of the proteins encoded by T4 genes 41, 45, 44, and 62 using a complementation assay.

作者信息

Nossal N G

出版信息

J Biol Chem. 1979 Jul 10;254(13):6026-31.

PMID:376524
Abstract

The proteins encoded by bacteriophage T4 genes 41, 45, 44, and 62 are known from the genetic studies of Epstein et al. ((1963) Cold Spring Harbor Symp. Quant. Biol. 28, 375--394) to be required for viral DNA synthesis. A convenient assay for each of these proteins is described which is based on the specific stimulation by each protein of DNA synthesis in extracts of Escherichia coli infected with mutants of bacteriophage T4 unable to make that protein. The T4 41 protein, 45 protein, and the complex of the 44 and 62 proteins have been highly purified. For each protein there is co-chromatography during the final purification step of (i) activity in the complementation assay, (ii) activity required for DNA synthesis with other purified T4 proteins, and (iii) a subunit of the size previously identified as that of the corresponding gene product.

摘要

通过爱泼斯坦等人((1963年)《冷泉港定量生物学研讨会》第28卷,第375 - 394页)的遗传学研究可知,噬菌体T4基因41、45、44和62编码的蛋白质是病毒DNA合成所必需的。本文描述了一种针对这些蛋白质中每一种的简便检测方法,该方法基于每种蛋白质对感染了无法产生该蛋白质的噬菌体T4突变体的大肠杆菌提取物中DNA合成的特异性刺激。T4 41蛋白、45蛋白以及44和62蛋白的复合物已被高度纯化。对于每种蛋白质,在最终纯化步骤中存在共色谱现象,具体表现为:(i)互补检测中的活性,(ii)与其他纯化的T4蛋白进行DNA合成所需的活性,以及(iii)先前鉴定为相应基因产物大小的一个亚基。

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