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伤寒杆菌Ty2的基因组切割图谱。

Genomic cleavage map of Salmonella typhi Ty2.

作者信息

Liu S L, Sanderson K E

机构信息

Department of Biological Sciences, University of Calgary, Alberta, Canada.

出版信息

J Bacteriol. 1995 Sep;177(17):5099-107. doi: 10.1128/jb.177.17.5099-5107.1995.

Abstract

The genomic cleavage map of Salmonella typhi Ty2, 4,780 kb in size, was determined through digestion of the genomic DNA with endonucleases and separation of the fragments by pulsed-field gel electrophoresis. The chromosome has 33, 26, 7, and 35 sites for the enzymes XbaI, BlnI, I-CeuI, and SpeI, respectively. The fragments were arranged around the chromosome through excision of fragments from the gel, redigestion with a second enzyme, and labelling with 32P, and reelectrophoresis and named in alphabetical order. Tn10 transposons inserted in 82 different genes of Salmonella typhimurium were transduced by phage P22 into S. typhi, and the location of Tn10, and thus of the gene, was mapped through the XbaI and BlnI sites of Tn10. All seven I-CeuI sites (in rrl genes for 23S rRNA) were conserved, and the gene order within the I-CeuI fragments resembles that of S. typhimurium LT2, but the order of I-CeuI fragments is rearranged from ABCDEFG in S. typhimurium LT2 to AGCEFDB in S. typhi. In addition, there is a 500-kb inversion which covers the terminus region. Comparisons of lengths of segments between genes showed that S. typhi has segments which differ in size from those in S. typhimurium. The viaB locus, for synthesis of the Vi antigen of S. typhi, was shown to be within a 118-kb loop (a segment of DNA with no homolog in most other Salmonella species) between mel and poxA on the chromosome.

摘要

伤寒杆菌Ty2的基因组切割图谱,大小为4780kb,是通过用核酸内切酶消化基因组DNA并通过脉冲场凝胶电泳分离片段来确定的。该染色体分别有33个、26个、7个和35个XbaI、BlnI、I-CeuI和SpeI酶切位点。通过从凝胶中切下片段、用第二种酶重新消化、用32P标记、再电泳,将片段围绕染色体排列,并按字母顺序命名。插入鼠伤寒沙门氏菌82个不同基因中的Tn10转座子被噬菌体P22转导到伤寒杆菌中,通过Tn10的XbaI和BlnI位点绘制了Tn10的位置,从而确定了基因的位置。所有七个I-CeuI位点(位于23S rRNA的rrl基因中)都是保守的,I-CeuI片段内的基因顺序与鼠伤寒沙门氏菌LT2相似,但I-CeuI片段的顺序从鼠伤寒沙门氏菌LT2中的ABCDEFG重排为伤寒杆菌中的AGCEFDB。此外,还有一个500kb的倒位,覆盖了末端区域。基因间片段长度的比较表明,伤寒杆菌的片段大小与鼠伤寒沙门氏菌不同。伤寒杆菌Vi抗原合成的viaB位点位于染色体上mel和poxA之间的一个118kb的环(一段在大多数其他沙门氏菌物种中无同源物的DNA)内。

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