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用于大肠杆菌K-12鞭毛基因(fla)的特异性λ转导噬菌体的分离

Isolation of specialized lambda transducing bacteriophages for flagellar genes (fla) of Escherichia coli K-12.

作者信息

Komeda Y, Shimada K, Iino T

出版信息

J Virol. 1977 Jun;22(3):654-61. doi: 10.1128/JVI.22.3.654-661.1977.

Abstract

Specialized transducing lambda phages carrying the region III flagellar genes (fla) of Escherichia coli K-12 were isolated by a new method. A strain carrying both a cryptic lambda prophage near the his genes and a deletion of the attlambda gene was used as a starting strain. The lysogen of lambdacI857pga18-bio69 was isolated in which the prophage was integrated within the lambda cryptic genes by means of recombination with the residual lambda DNA. The strains with deletions starting within the prophage and ending in these fla genes were selected from among the heat-resistant survivors of the lysogen. They were then infected with heat-inducible and lysis-defective lambda phages and, thus, specialized transducing phage lines for hag and fla were obtained. High-frequency transfer lines of rare phages carrying the fla genes were isolated by inducing a strain carrying a heat-inducible lambda prophage near the his genes and selecting by transduction of a fla deletion strain. Preliminary characterization of these transducing phages is also reported.

摘要

通过一种新方法分离出了携带大肠杆菌K-12 III区鞭毛基因(fla)的特异性转导λ噬菌体。使用在his基因附近携带一个隐蔽λ原噬菌体且attlambda基因缺失的菌株作为起始菌株。分离出λcI857pga18-bio69的溶原菌,其中原噬菌体通过与残留的λDNA重组整合到λ隐蔽基因内。从溶原菌的耐热存活菌中筛选出从原噬菌体内部开始缺失并在这些fla基因处结束的菌株。然后用热诱导且裂解缺陷的λ噬菌体感染它们,从而获得了针对hag和fla的特异性转导噬菌体株系。通过诱导在his基因附近携带热诱导λ原噬菌体的菌株并通过转导fla缺失菌株进行筛选,分离出了携带fla基因的稀有噬菌体的高频转移株系。还报道了这些转导噬菌体的初步特征。

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