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大肠杆菌K-12中大肠杆菌素E1 DNA的特异性转导

Specialized transduction of colicin E1 DNA in Escherichia coli K-12.

作者信息

Fukumaki Y, Shimada K, Takagi Y

出版信息

Proc Natl Acad Sci U S A. 1976 Sep;73(9):3238-42. doi: 10.1073/pnas.73.9.3238.

Abstract

Genetic studies were made on E. coli K-12 TM96, which carries recombinant molecules constructed by in vitro combination of colicin E1 DNA and a DNA fragment of E. coli for guanine synthesis derived from transducing phage. The recombinant molecules existed as stable plasmids within the cell and contained genes for colicin E1 immunity and the guaA enzyme (xanthosine 5'-monophosphate aminase) together with a part of the lambda genome, R through J: (R-A-F-J)+. A block of the lambda genome, int through Q, was not detected in the recombinant molecule. Thus, this recombinant molecule was named ColEl-coslambda-guaA, and the specialized tranduction of the ColEl-coslambda-guA DNA into various E. coli K-12 cells by lambda phage was described. Lysates prepared by lytic infection of lambda phage onto TM96 or by induction of TM96(lambda) lysogens contained transducing particles which could transduce gua-deleted E. coli to stable guaA+ cells. These transductants were proved to have similar genetic properties as those of TM96. The frequency of transduction was not affected by the presence of an attachement site for lambda, prophage lambda, colicin E1 plasmids, or the recA property within gua-deleted recipient cells. Transducing particles were resistant to EDTA treatment and most of them had an average density of about 1.472. This value corresponds to that of lambda phage particles, which contain about 72% of the lenght of lambda DNA.

摘要

对大肠杆菌K-12 TM96进行了遗传学研究,该菌株携带通过体外组合大肠杆菌素E1 DNA和来自转导噬菌体的用于鸟嘌呤合成的大肠杆菌DNA片段构建的重组分子。重组分子在细胞内以稳定质粒的形式存在,包含大肠杆菌素E1免疫基因和guaA酶(黄苷5'-单磷酸氨基酶)以及λ基因组的一部分,从R到J:(R-A-F-J)+。在重组分子中未检测到λ基因组的一段,即从int到Q。因此,这种重组分子被命名为ColEl-cosλ-guaA,并描述了通过λ噬菌体将ColEl-cosλ-guA DNA特异性转导到各种大肠杆菌K-12细胞中的过程。通过λ噬菌体对TM96进行裂解感染或诱导TM96(λ)溶原菌制备的裂解物含有转导颗粒,这些颗粒可以将gua缺失的大肠杆菌转导为稳定的guaA+细胞。这些转导子被证明具有与TM96相似的遗传特性。转导频率不受gua缺失的受体细胞中λ附着位点、原噬菌体λ、大肠杆菌素E1质粒或recA特性的影响。转导颗粒对EDTA处理具有抗性,并且它们中的大多数平均密度约为1.472。该值与λ噬菌体颗粒的值相对应,λ噬菌体颗粒包含约72%的λDNA长度。

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引用本文的文献

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本文引用的文献

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Dilysogenic excision: an accessory expression of the termination function?
Cold Spring Harb Symp Quant Biol. 1968;33:749-54. doi: 10.1101/sqb.1968.033.01.085.
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The integration and excision of the bacteriophage lambda genome.噬菌体λ基因组的整合与切除
Cold Spring Harb Symp Quant Biol. 1968;33:735-47. doi: 10.1101/sqb.1968.033.01.084.
9
Replication of colicin E1 plasmid DNA in cell extracts.大肠杆菌素E1质粒DNA在细胞提取物中的复制
Proc Natl Acad Sci U S A. 1974 Mar;71(3):802-6. doi: 10.1073/pnas.71.3.802.

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