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噬菌斑形成的λdnaZ⁺转导噬菌体的分离与鉴定

Isolation and characterization of plaque-forming lambdadnaZ+ transducing bacteriophages.

作者信息

Walker J R, Henson J M, Lee C S

出版信息

J Bacteriol. 1977 Apr;130(1):354-65. doi: 10.1128/jb.130.1.354-365.1977.

Abstract

The Escherichia coli dnaZ gene, a deoxyribonucleic acid (DNA) polymerization gene, is located 1.2 min counterclockwise from purE, at approximately min 10.5 on the E. coli map. From a lysogen with lamdacI857 integrated at a secondary attachment site near purE, transducing phages (lambdadnaS+) that transduced a dnaZts (lambda+) recipient to temperature insensitivity (TS+) were discovered. Three different plaque-forming transducing phages were isolated from seven primary heterogenotes. Genetic tests and heteroduplex mapping were used to determine the length and position of E. coli DNA within the lambda DNA. Complementation tests demonstrated that the deletions in all three strains removed both att P and the int gene, i,e., DNA from both prophage ends. Heteroduplex mapping confirmed this result by demonstrating that all three strains had deletions of lambda DNA that covered the b2 to red region, thereby removing both prophage ends. Specifically, the deletions removed lambda DNA between the points 39.3 to 66.5% of lambda length (measured in percent length from the left and of lambda phage DNA) in all three strains. The three strains are distinct, however, because they had differing lengths of host DNA insertions. These phages must have been formed by an anomalous procedure, because standard lambda transducing phages are deleted for one prophage end only. In lambdagal and lambdabio strains, the deletions of lambda DNA begin at the union of prophage ends (i.e., position 57.3% of lambda length) and extend leftward or rightward, respectively (Davidson and Szybalski, in A, D. Hershey [ed.], The Bacteriophage Lambda, p. 45-82, 1971). Models for formation of the lambdadnaZ+ phages are discussed.

摘要

大肠杆菌dnaZ基因是一种脱氧核糖核酸(DNA)聚合基因,位于purE逆时针方向1.2分钟处,在大肠杆菌染色体图上约为10.5分钟处。从一个原噬菌体lamdacI857整合在靠近purE的二级附着位点的溶原菌中,发现了能将dnaZts(lambda+)受体转导为温度不敏感型(TS+)的转导噬菌体(lambdadnaS+)。从七个初级杂合子中分离出三种不同的噬菌斑形成转导噬菌体。通过遗传测试和异源双链体图谱分析来确定lambda DNA内大肠杆菌DNA的长度和位置。互补测试表明,所有三个菌株中的缺失都去除了att P和int基因,即来自原噬菌体两端的DNA。异源双链体图谱分析证实了这一结果,表明所有三个菌株的lambda DNA缺失都覆盖了b2到red区域,从而去除了原噬菌体两端。具体而言,所有三个菌株中的缺失都去除了lambda长度39.3%至66.5%之间的lambda DNA(从lambda噬菌体DNA的左端开始以长度百分比测量)。然而,这三个菌株是不同的,因为它们的宿主DNA插入片段长度不同。这些噬菌体肯定是通过异常过程形成的,因为标准的lambda转导噬菌体只缺失一个原噬菌体末端。在lambdagal和lambdabio菌株中,lambda DNA的缺失分别从原噬菌体两端的连接处(即lambda长度的57.3%处)开始向左或向右延伸(戴维森和齐巴尔斯基,载于A.D. 赫希编《噬菌体lambda》,第45 - 82页,1971年)。文中讨论了lambdadnaZ+噬菌体的形成模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9fb7/235213/3078b6b3a828/jbacter00305-0373-a.jpg

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