Defined single-gene and multi-gene deletion mutant collections in salmonella enterica sv typhimurium

Steffen Porwollik, Carlos A. Santiviago, Pui Cheng, Fred Long, Prerak Desai, Jennifer Fredlund, Shabarinath Srikumar, Cecilia A. Silva, Weiping Chu, Xin Chen, Rocío Canals, M. Megan Reynolds, Lydia Bogomolnaya, Christine Shields, Ping Cui, Jinbai Guo, Yi Zheng, Tiana Endicott-Yazdani, Hee Jeong Yang, Aimee MapleYury Ragoza, Carlos J. Blondel, Camila Valenzuela, Helene Andrews-Polymenis, Michael McClelland

Research output: Contribution to journalArticlepeer-review

118 Citations (Scopus)

Abstract

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 (Cam R) 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 (www.beiresources.org) and publicly available.

Original languageEnglish
Article numbere99820
JournalPLoS ONE
Volume9
Issue number7
DOIs
Publication statusPublished - 9 Jul 2014

ASJC Scopus subject areas

  • General Biochemistry,Genetics and Molecular Biology
  • General Agricultural and Biological Sciences

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