Chromosomal Complementation Using Tn7 Transposon Vectors in Enterobacteriaceae
Chromosomal Complementation Using Tn7 Transposon Vectors in Enterobacteriaceae
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DOI：
10.1128/aem.00986-12
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发表时间：
2012-09-01
影响因子：
4.4
通讯作者：
Dozois, Charles M.
中科院分区：
文献类型：
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作者：
Crepin, Sebastien;Harel, Jos;Dozois, Charles M.
许多细菌中的遗传互补通常通过在重组质粒上重新引入突变或缺失基因的功能拷贝来实现。使用Tn 7转座子的染色体整合系统具有提供不需要选择压力的稳定单拷贝整合的优点。以前的Tn 7系统已经被开发出来，尽管没有一个被证明在各种肠细菌中有效地工作。我们已经开发了几种mini-Tn 7和转座酶载体，以提供更通用的系统。Tn 7在染色体attTn 7位点的转座通过经典的接合方法实现，其中供体菌株携带mini-Tn 7载体，受体菌株具有转座酶载体。这种方法对五种不同的致病性肠细菌是有效的。因此，该系统提供了一个有用的工具，在一个游离位点的单拷贝互补的细菌遗传学和微生物的发病机制的研究。此外，这些载体还可用于引入外源基因，用于生物技术应用、疫苗开发或基因表达和基因融合构建体。
Genetic complementation in many bacteria is commonly achieved by reintroducing functional copies of the mutated or deleted genes on a recombinant plasmid. Chromosomal integration systems using the Tn7 transposon have the advantage of providing a stable single-copy integration that does not require selective pressure. Previous Tn7 systems have been developed, although none have been shown to work effectively in a variety of enterobacteria. We have developed several mini-Tn7 and transposase vectors to provide a more versatile system. Transposition of Tn7 at the chromosomal attTn7 site was achieved by a classical conjugation approach, wherein the donor strain harbored the mini-Tn7 vector and the recipient strain possessed the transposase vector. This approach was efficient for five different pathogenic enterobacterial species. Thus, this system provides a useful tool for single-copy complementation at an episomal site for research in bacterial genetics and microbial pathogenesis. Furthermore, these vectors could also be used for the introduction of foreign genes for use in biotechnology applications, vaccine development, or gene expression and gene fusion constructs.