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缺陷噬菌体15所含脱氧核糖核酸的特性。

Properties of the deoxyribonucleic acid contained in the defective particle coliphage 15.

作者信息

Frampton E W, Mandel M

出版信息

J Virol. 1970 Jan;5(1):8-13. doi: 10.1128/JVI.5.1.8-13.1970.

Abstract

Escherichia coli strain 15 TAU, which requires thymine, arginine, and uracil for growth and harbors an apparently defective prophage, was induced by exposure to ultraviolet light (580 ergs/mm(2)) or to mitomycin C (5 mug/ml). Phage particles (coliphage 15) were recovered from the resulting lysate by treatment with deoxyribonuclease, filtration, and several cycles of differential centrifugation. Analysis of the phage particles obtained by using cesium chloride density gradient centrifugation in a preparative ultracentrifuge resulted in the resolution of three components. The major component had a peak density of 1.52 to 1.53 g/cm(3) followed by components with densities of 1.5 and 1.49 g/cm(3). The guanine plus cytosine content of coliphage 15 deoxyribonucleic acid (DNA) was determined by both analytical ultracentrifugation in cesium chloride and by thermal denaturation in standard saline citrate buffer. Respective values of 46.4 +/- 1% and 46.6 +/- 1% guanine plus cytosine content were obtained. Coliphage 15 DNA formed molecular hybrids with messenger ribonucleic acid (RNA) from both uninduced and ultraviolet-induced cultures of E. coli 15 TAU, but did not hybridize with E. coli ribosomal RNA. The molecular weight of coliphage 15 DNA was determined by constant velocity sucrose density gradient centrifugation to be about 33 x 10(6) daltons.

摘要

大肠杆菌15 TAU菌株生长需要胸腺嘧啶、精氨酸和尿嘧啶,且带有一个明显有缺陷的原噬菌体,通过紫外线(580尔格/平方毫米)或丝裂霉素C(5微克/毫升)处理可诱导其产生噬菌体。用脱氧核糖核酸酶处理、过滤以及多次差速离心后,从所得裂解物中回收噬菌体颗粒(大肠杆菌噬菌体15)。在制备型超速离心机中使用氯化铯密度梯度离心法对获得的噬菌体颗粒进行分析,结果分离出三个组分。主要组分的峰值密度为1.52至1.53克/立方厘米,随后是密度为1.5和1.49克/立方厘米的组分。通过在氯化铯中进行分析超速离心以及在标准柠檬酸盐缓冲盐溶液中进行热变性,测定了大肠杆菌噬菌体15脱氧核糖核酸(DNA)中鸟嘌呤加胞嘧啶的含量。鸟嘌呤加胞嘧啶含量的相应值分别为46.4±1%和46.6±1%。大肠杆菌噬菌体15 DNA与来自未诱导和紫外线诱导的大肠杆菌15 TAU培养物的信使核糖核酸(RNA)形成分子杂交体,但不与大肠杆菌核糖体RNA杂交。通过等速蔗糖密度梯度离心法测定大肠杆菌噬菌体15 DNA的分子量约为33×10⁶道尔顿。

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