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λ转导噬菌体以及携带大肠杆菌K12的cysJIHDC基因簇基因的克隆

Lambda transducing phage and clones carrying genes of the cysJIHDC gene cluster of Escherichia coli K12.

作者信息

Hunt C L, Colless V, Smith M T, Molasky D O, Malo M S, Loughlin R E

机构信息

Department of Biochemistry, University of Sydney, New South Wales, Australia.

出版信息

J Gen Microbiol. 1987 Oct;133(10):2707-17. doi: 10.1099/00221287-133-10-2707.

Abstract

DNA from each of two specialized transducing lambda phage, lambda dcysJIHD and lambda cysJ, has been analysed by heteroduplex mapping. The segment of the Escherichia coli chromosome carried by lambda dcysJIHD was shown to be large, approximately 18 kb in length, and to replace a large length of lambda DNA, approximately 11 kb, which includes the genes for integration and recombination. Thus lambda dcysJIHD is a bio-type transducing phage. lambda cysJ was shown to have lost very little lambda DNA and to carry about 8 kb of bacterial DNA. Sites for several restriction endonucleases were mapped in the DNA from each phage and cloning experiments located some of the genes of the cluster in relation to the restriction map. Cysteine regulation of the cloned cysJ and cysD genes was shown as well as cysteine regulation of beta-galactosidase in some constructs. The direction of transcription of the cysD gene was established, and from physical evidence the size of the 'silent section' between the cysH and cysD genes was estimated to be at least 11 kb.

摘要

已通过异源双链图谱分析了两种特异性转导λ噬菌体(λdcysJIHD和λcysJ)的DNA。结果表明,λdcysJIHD携带的大肠杆菌染色体片段很大,长度约为18 kb,并取代了约11 kb的大片段λDNA,其中包括整合和重组基因。因此,λdcysJIHD是一种生物型转导噬菌体。λcysJ显示出丢失的λDNA很少,并携带约8 kb的细菌DNA。在每个噬菌体的DNA中绘制了几种限制性内切酶的位点,克隆实验确定了该基因簇的一些基因与限制性图谱的关系。结果显示了克隆的cysJ和cysD基因的半胱氨酸调节以及某些构建体中β-半乳糖苷酶的半胱氨酸调节。确定了cysD基因的转录方向,根据物理证据估计cysH和cysD基因之间“沉默区”的大小至少为11 kb。

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