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球形红假单胞菌噬菌体RS1 DNA的结构与物理图谱

Structure and physical map of Rhodopseudomonas sphaeroides bacteriophage RS1 DNA.

作者信息

Donohue T J, Chory J, Goldsand T E, Lynn S P, Kaplan S

出版信息

J Virol. 1985 Jul;55(1):147-57. doi: 10.1128/JVI.55.1.147-157.1985.

Abstract

We analyzed, by restriction endonuclease mapping and electron microscopy, the genome of the lytic Rhodopseudomonas sphaeroides-specific bacteriophage RS1 and characterized it as a linear molecule of approximately 60 to 65 kilobases. When the DNA from purified phage particles was examined by several independent methods, considerable size heterogeneity was apparent in the RS1 DNA. This size heterogeneity was concluded to be of biological origin, was independent of the specific host strain used to propagate virus, and was not due to the presence of host DNA within or nonspecifically associated with purified virions. In addition, treatment of RS1 DNA with either BAL 31 nuclease or DNA polymerase I Klenow fragment revealed that several distinct regions exist within the viral chromosome which contain free 3' hydroxyl groups. A restriction endonuclease map of the RS1 genome was constructed by using the restriction endonucleases EcoRI, ClaI, KpnI, BamHI, MluI, SmaI, and BclI; thereby allowing the positioning of some 40 restriction sites within the viral genome. The results are discussed in terms of the significance and the possible biological origin of the unique features discovered within the phage RS1 DNA.

摘要

我们通过限制性内切酶图谱分析和电子显微镜技术,对溶菌性球形红假单胞菌特异性噬菌体RS1的基因组进行了分析,并将其鉴定为一个大小约为60至65千碱基的线性分子。当用几种独立方法检测纯化噬菌体颗粒的DNA时,RS1 DNA中明显存在相当大的大小异质性。这种大小异质性被认为源于生物学因素,与用于繁殖病毒的特定宿主菌株无关,也不是由于纯化病毒粒子内部存在宿主DNA或与宿主DNA非特异性结合所致。此外,用BAL 31核酸酶或DNA聚合酶I Klenow片段处理RS1 DNA后发现,病毒染色体中存在几个不同区域,这些区域含有游离的3'羟基。利用限制性内切酶EcoRI、ClaI、KpnI、BamHI、MluI、SmaI和BclI构建了RS1基因组的限制性内切酶图谱,从而确定了病毒基因组内约40个限制性位点的位置。我们根据在噬菌体RS1 DNA中发现的独特特征的意义和可能的生物学来源对结果进行了讨论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32b9/254909/65df39bfd365/jvirol00118-0159-a.jpg

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