Directed formation of chromosomal deletions in Salmonella typhimurium: targeting of specific genes induced during infection.
Directed formation of chromosomal deletions in Salmonella typhimurium: targeting of specific genes induced during infection.
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鼠伤寒沙门氏菌染色体缺失的定向形成：针对感染期间诱导的特定基因。
DOI：
10.1007/s004380050721
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Mahan,MJ
中科院分区：
文献类型：
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作者：
Julio,SM;Conner,CP;Heithoff,DM;Mahan,MJ
体内表达技术 (IVET) 已分离出 100 多种在感染过程中诱导产生的鼠伤寒沙门氏菌。许多体内诱导 (ivi) 基因以及其他毒力基因聚集在沙门氏菌特异的染色体区域，而大肠杆菌中不存在（例如致病岛）。希望能够删除染色体的这种假定的毒力区域，并且如果删除删除了在发病机制中发挥作用的基因，则随后的努力可以集中于驻留在该区域内的单个基因。因此，我们开发了一种构建染色体缺失的策略，其大小不受限制，在连接点处用可选择标记定义了终点，并且不依赖于删除区域内包含的序列的先验知识。这种删除策略可以应用于几乎所有具有同源重组的细菌以及不需要或不可行同源重组的基于质粒的突变系统。
In vivo expression technology (IVET) has resulted in the isolation of more than 100Salmonella typhimuriumgenes that are induced during infection. Many of these in vivo induced (ivi) genes, as well as other virulence genes, are clustered in regions of the chromosome that are specific forSalmonellaand are not present inEscherichia coli(e.g., pathogenicity islands). It would be desirable to be able to delete such putative virulence regions of the chromosome, and if the deletion removes genes that play a role in pathogenesis subsequent efforts can then be focused on individual genes that reside within that region. We therefore have developed a strategy for constructing chromosomal deletions which are not limited in size, have defined endpoints with a selectable marker at the joint point, and are not dependent on prior knowledge of sequences contained within the deleted region. Such deletion strategies can be applied to almost any bacterium with homologous recombination and to plasmid-based mutational systems where homologous recombination is not desired or feasible.