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体外phiX174依赖DNA的DNA合成：大肠杆菌dnaB突变提取物中P1禁令蛋白的需求

phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.

作者信息

Schuster H, Mikolajczyk M, Rohrschneider J, Geschke B

出版信息

Proc Natl Acad Sci U S A. 1975 Oct;72(10):3907-11. doi: 10.1073/pnas.72.10.3907.

DOI:10.1073/pnas.72.10.3907

PMID:670

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC433105/

Abstract

Ammonium sulfate fractionation of crude extracts of E. coli yields a soluble enzyme fraction (about 25-fold purification) that catalyzes the conversion of phiX174 single-stranded DNA to duplex DNA. The reaction is rifampicin-resistant, requires single-stranded DNA, Mg++, deoxynucleoside triphosphates, and ATP, and is stimulated by KCl. Such soluble enzyme fractions were prepared from E. coli strains carrying the prophage mutant P1bac, in which the viral dnaB analog (ban) protein is expressed constitutively, or P1bacban, in which the expression of ban protein is prevented. DNA-synthesizing activity of ban protein containing fractions from wild-type or dnaB(P1bac) lysogens was more temperature-resistant than that from E. coli containing only wild-type dnaB protein, whereas that from dnaB(P1bacban) lysogens of dnaB cells was extremely thermolabile. It is suggested that the temperature-resistant DNA synthesis with fractions from P1bac lysogens is mediated by the P1 ban protein.

摘要

用硫酸铵对大肠杆菌粗提物进行分级分离，可得到一种可溶性酶级分（约25倍纯化），该酶级分催化φX174单链DNA转化为双链DNA。该反应对利福平具有抗性，需要单链DNA、Mg++、脱氧核苷三磷酸和ATP，并受KCl刺激。这种可溶性酶级分是从携带原噬菌体突变体P1bac的大肠杆菌菌株制备的，其中病毒dnaB类似物（ban）蛋白组成性表达，或从P1bacban制备的，其中ban蛋白的表达被阻止。来自野生型或dnaB（P1bac）溶原菌的含ban蛋白级分的DNA合成活性比仅含野生型dnaB蛋白的大肠杆菌的DNA合成活性更耐温度，而来自dnaB细胞的dnaB（P1bacban）溶原菌的DNA合成活性则极其不耐热。提示来自P1bac溶原菌级分的耐温度DNA合成是由P1 ban蛋白介导的。