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携带metBJF甲硫氨酸基因簇的特异性λ转导噬菌体的分离与鉴定

Isolation and characterization of specialized lambda transducing bacteriophage carrying the metBJF methionine gene cluster.

作者信息

Johnson J R, Greene R C, Krueger J H

出版信息

J Bacteriol. 1977 Sep;131(3):795-800. doi: 10.1128/jb.131.3.795-800.1977.

Abstract

Secondary attachment site lysogens of Deltaatt(lambda)Deltappc-argECBH strains of Escherichia coli with lambdacI857 integrated into the bfe gene (88 min) were isolated. Of 20 such lysogens examined, 2 produce lysates with transducing phage containing the metBJF gene cluster (87 min). Reintroduction of the ppc-argECBH chromosome segment (which lies between the bfe and met genes) into these strains virtually abolishes the production of met transducing phage. All of the phage examined have lost essential genes from the left arm of the lambda chromosome. Approximately 85% of the phage appear to have the same genetic composition, containing the metBJF gene cluster, but not the closely linked gene cytR, and having lost phage genes G and J. Analytical CsCl density gradient centrifugation of five representatives of this major class of phage shows four of them to have identical densities (lighter than lambda), while the fifth cannot be resolved from lambda. The four apparently identical phage were isolated from three separate lysates, which suggests the existence of preferred sites for illegitimate recombination on the bacterial and phage chromosomes. Three specialized transducing phage that carry cytR in addition to metB, metJ, and metF have also been studied. Each of these viruses has a different amount of phage deoxyribonucleic acid. Two of them have less deoxyribonucleic acid than lambda, whereas the third has about the same amount. The metB, metF, and cytR genes of the transducing phage have been shown to function in vivo. The phage-borne metB and metF genes are subject to metJ-mediated repression.

摘要

分离出了大肠杆菌Deltaatt(lambda)Deltappc - argECBH菌株的二级附着位点溶原菌,其中lambda cI857整合到了bfe基因(88分钟处)。在检测的20株这样的溶原菌中,有2株产生的裂解物带有含有metBJF基因簇(87分钟处)的转导噬菌体。将ppc - argECBH染色体片段(位于bfe和met基因之间)重新导入这些菌株后,几乎完全消除了met转导噬菌体的产生。所有检测的噬菌体都丢失了lambda染色体左臂上的必需基因。大约85%的噬菌体似乎具有相同的基因组成,包含metBJF基因簇,但不包含紧密连锁的基因cytR,并且丢失了噬菌体基因G和J。对这一主要类型噬菌体的五个代表进行分析性CsCl密度梯度离心,结果显示其中四个具有相同的密度(比lambda轻),而第五个无法与lambda区分开来。这四个明显相同的噬菌体是从三个不同的裂解物中分离出来的,这表明在细菌和噬菌体染色体上存在非法重组的优先位点。还研究了三种除了携带metB、metJ和metF之外还携带cytR的特异性转导噬菌体。这些病毒中的每一种都有不同量的噬菌体脱氧核糖核酸。其中两种的脱氧核糖核酸比lambda少,而第三种的量与lambda大致相同。已证明转导噬菌体的metB、metF和cytR基因在体内发挥功能。噬菌体携带的metB和metF基因受到metJ介导的抑制。

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