PLoS ONE (Jan 2014)

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 Maple,
  • Yury Ragoza,
  • Carlos J Blondel,
  • Camila Valenzuela,
  • Helene Andrews-Polymenis,
  • Michael McClelland

DOI
https://doi.org/10.1371/journal.pone.0099820
Journal volume & issue
Vol. 9, no. 7
p. e99820

Abstract

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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 (www.beiresources.org) and publicly available.