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携带从异常原噬菌体插入F因子的traY基因中分离出的转移基因的λ转导噬菌体。

lambda-Transducing phages carrying transfer genes isolated from an abnormal prophage insertion into the traY gene of F.

作者信息

Johnson D A, Willetts N S

出版信息

Plasmid. 1983 Jan;9(1):71-85. doi: 10.1016/0147-619x(83)90032-x.

Abstract

A set of lambda-transducing phages carrying transfer (tra) genes has been isolated from an abnormal lysogen in which a lambda prophage was inserted into the traY gene of Flac. These have been characterized genetically for complementation of Flac tra and finP point mutants and for the presence of oriT. Studies of tra gene expression during lambda repression showed that tra genes on the transducing phages were expressed from the lambda PL promoter as well as from the transfer promoters when these were present. The molecular weights of the traM (14,000) and traJ (23,500) proteins were measured after infection of ultraviolet-irradiated cells with one of the phages, ED lambda 102, and overproduction of the traJ protein upon induction of an ED lambda 102 lysogen was demonstrated. A proportion of this traJ protein was located in the inner membrane and cytoplasmic fractions of the cell, the majority being in the outer membrane. Physical analysis of the DNA carried by the lambda tra phages by determination of the phage buoyant densities in CsCl, by restriction enzyme digestion and by electron microscope heteroduplex analysis, was used to define the DNA segments encoding the tra functions. Correlation of the physical and genetical data improved the positioning of the tra genes within the transfer region. These results were combined with new restriction enzyme cleavage data to construct an improved map of this region.

摘要

一组携带转移(tra)基因的λ转导噬菌体已从一个异常溶原菌中分离出来,在该溶原菌中,一个λ原噬菌体插入到Flac的traY基因中。这些噬菌体已通过对Flac tra和finP点突变体的互补作用以及oriT的存在进行了遗传学表征。对λ抑制期间tra基因表达的研究表明,当存在转移启动子时,转导噬菌体上的tra基因可从λ PL启动子以及转移启动子表达。在用其中一种噬菌体EDλ102感染紫外线照射的细胞后,测定了traM(14,000)和traJ(23,500)蛋白的分子量,并证明了诱导EDλ102溶原菌时traJ蛋白的过量产生。该traJ蛋白的一部分位于细胞的内膜和细胞质部分,大部分位于外膜。通过测定噬菌体在CsCl中的浮力密度、用限制酶消化以及通过电子显微镜异源双链分析对λ tra噬菌体携带的DNA进行物理分析,以确定编码tra功能的DNA片段。物理和遗传数据的相关性改善了tra基因在转移区域内的定位。这些结果与新的限制酶切割数据相结合,构建了该区域的改进图谱。

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