A self-deleting Cre-lox-ermAM cassette, Cheshire, for marker-less gene deletion in Streptococcus pneumoniae.
A self-deleting Cre-lox-ermAM cassette, Cheshire, for marker-less gene deletion in Streptococcus pneumoniae.
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DOI：
10.1016/j.mimet.2009.10.007
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发表时间：
2009-12
影响因子：
2.2
通讯作者：
Morrison, Donald A.
中科院分区：
文献类型：
--
作者：
Weng, Liming;Biswas, Indranil;Morrison, Donald A.
尽管肺炎链球菌的靶向诱变在天然遗传转化和体外组装的嵌合供体DNA构建体的帮助下容易地完成，但通常用于选择重组产物的耐药标记物本身可能是遗传操作的不期望的副产物。描述了携带红霉素抗性标记ermAM的新盒，其可用作用于选择所需重组体的临时标记。随后可以通过嵌入的岩藻糖调节的Cre重组酶基因和末端lox66和lox71 Cre识别位点随意去除盒，保留盒中的34 bp作为惰性残留双突变体lox72位点。
Although targeted mutagenesis of Streptococcus pneumoniae is readily accomplished with the aid of natural genetic transformation and chimeric donor DNA constructs assembled in vitro, the drug resistance markers often employed for selection of recombinant products can themselves be undesirable by-products of the genetic manipulation. A new cassette carrying the erythromycin-resistance marker ermAM is described that can be used as a temporary marker for selection of desired recombinants. The cassette may subsequently be removed at will by virtue of an embedded fucose-regulated Cre recombinase gene and terminal lox66 and lox71 Cre recognition sites, with retention of 34 bp from the cassette as an inert residual double-mutant lox72 site.
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