Plasmids for ectopic integration in Bacillus subtilis
Plasmids for ectopic integration in Bacillus subtilis
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DOI：
10.1016/s0378-1119(96)00404-0
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发表时间：
1996-11-21
期刊：
影响因子：
3.5
通讯作者：
Stragier, P
中科院分区：
文献类型：
--
作者：
GueroutFleury, AM;Frandsen, N;Stragier, P
已经构建的质粒允许在枯草芽孢杆菌(BS)染色体的thC基因座上通过双重组事件进行整合。这些质粒既可用于构建分二倍体菌株和互补分析，也可用于构建与大肠杆菌LacZ基因的转录融合。该质粒含有一个可在BS中选择的抗生素(AN)标记，以及一个额外的可重组到染色体中的区域外的标记。当与ThrC位点含有第三个AN标记的受体菌株一起使用时，这些质粒可以容易地识别从标记交换事件中发出的转化子，而不需要额外的Campbell类型整合。在Amye基因座用于异位整合的现有质粒也进行了类似的修改。
Plasmids have been constructed that allow integration by a double recombination event at the thrC locus of the Bacillus subtilis (Bs) chromosome. These plasmids can be used either for construction of merodiploid strains and complementation analysis, or for construction of transcriptional fusions to the Escherichia coli lacZ gene. The plasmids contain an antibiotic (An) marker selectable in Bs, as well as an additional An marker outside of the region that can recombine into the chromosome. When used in conjunction with recipient strains containing a third An marker at their thrC locus, these plasmids allow easy identification of transformants issued from a marker exchange event without additional Campbell-type integration. The existing plasmids used for ectopic integration at the amyE locus have been modified similarly.