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Defined Single-Gene and Multi-Gene Deletion Mutant Collections in Salmonella enterica sv Typhimurium

机译:鼠伤寒沙门氏菌鼠伤寒沙门氏菌中定义的单基因和多基因缺失突变体集合

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摘要

We constructed two collections of targeted single gene deletion (SGD) mutants and two collections of targeted multi-gene deletion (MGD) mutants in Salmonella enterica sv Typhimurium 14028s. The SGD mutant collections contain (1), 3517 mutants in which a single gene is replaced by a cassette containing a kanamycin resistance (KanR) gene oriented in the sense direction (SGD-K), and (2), 3376 mutants with a chloramphenicol resistance gene (CamR) oriented in the antisense direction (SGD-C). A combined total of 3773 individual genes were deleted across these SGD collections. The MGD collections contain mutants bearing deletions of contiguous regions of three or more genes and include (3), 198 mutants spanning 2543 genes replaced by a KanR cassette (MGD-K), and (4), 251 mutants spanning 2799 genes replaced by a CamR cassette (MGD-C). Overall, 3476 genes were deleted in at least one MGD collection. The collections with different antibiotic markers permit construction of all viable combinations of mutants in the same background. Together, the libraries allow hierarchical screening of MGDs for different phenotypic followed by screening of SGDs within the target MGD regions. The mutants of these collections are stored at BEI Resources () and publicly available.
机译:我们在鼠伤寒沙门氏菌鼠伤寒沙门氏菌14028s中构建了两个针对性的单基因缺失(SGD)突变体和两个针对性的多基因缺失(MGD)突变体。 SGD突变体集合包含(1)个,3517个突变体,其中单个基因被包含一个沿有义方向(SGD-K)定向的卡那霉素抗性(Kan R )基因的盒替代,以及( 2),有3376个突变体,其反方向带有氯霉素抗性基因(Cam R )(SGD-C)。这些SGD集合中总共删除了3773个个体基因。 MGD集合包含携带三个或更多基因连续区域缺失的突变体,包括(3),横跨2543个基因的198个突变体,被Kan R 盒(MGD-K)取代,以及(4), Cam R 盒(MGD-C)取代了251个跨越2799个基因的突变体。总体而言,至少一个MGD集合中删除了3476个基因。具有不同抗生素标记的集合允许在相同背景下构建突变体的所有可行组合。这些库一起允许对不同表型的MGD进行分级筛选,然后在目标MGD区域内筛选SGD。这些集合的突变体存储在BEI Resources()中并公开可用。

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