Broad-host-range cre-lox system for antibiotic marker recycling in Gram-negative bacteria
Broad-host-range cre-lox system for antibiotic marker recycling in Gram-negative bacteria
复制标题
DOI：
10.2144/02335rr01
复制
发表时间：
2002-11-01
期刊：
影响因子：
2.7
通讯作者：
Lidstrom, ME
中科院分区：
文献类型：
--
作者：
Marx, CJ;Lidstrom, ME
现在，许多缺乏复杂遗传工具的细菌物种的完整基因组序列都是可用的。我们描述了一个广泛的主机范围crelox系统，允许在各种革兰氏阴性菌的抗生素标记回收的发展。该系统由等位基因交换载体和四环素抗性IncP质粒组成，所述等位基因交换载体携带侧翼为loxP位点的卡那霉素盒，所述四环素抗性IncP质粒提供Cre重组酶的表达。我们通过在两种不同的细菌，扭脱甲基杆菌AM1和真菌伯克霍尔德氏菌LB 400中产生未标记的基因缺失来证明该系统。这种新的抗生素标记回收系统提供了在各种革兰氏阴性细菌中产生未标记突变体的可能性。此外，标记再循环允许在目前几乎没有抗生素标记可用的生物体中产生携带多种遗传操作的菌株。
Complete genome sequences are now available for many bacterial species that lack sophisticated genetic tools. We describe the development of a broad-host-range crelox system that allows antibiotic marker recycling in a variety of Gram-negative bacteria. This system consists of an allelic exchange vector bearing a kanamycin cassette flanked by loxP sites and a tetracycline-resistant IncP plasmid that provides expression of the Cre recombinase. We demonstrate this system by generating unmarked deletions of genes in two different bacteria, Methylobacterium extorquens AM1 and Burkholderia fungorum LB400. This new antibiotic marker recycling system offers the possibility of creating unmarked mutants in a wide variety, of Gram-negative bacteria. Furthermore, marker recycling allows the generation of strains bearing multiple genetic manipulations in organisms for which few antibiotic markers are currently available.