Construction of targeted insertion mutations in Francisella tularensis subsp. novicida

Date
2007-10
Authors
Liu, Jirong
Zogaj, Xhavit
Barker, Jeffrey R.
Klose, Karl E.
Journal Title
Journal ISSN
Volume Title
Publisher
Future Science Group
Abstract

Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance cassettes and splicing by overlap extension (SOE). This process enables fast and efficient construction of targeted insertion mutations in F. tularensis subsp. novicida that have characteristics of nonpolar mutations; optimized targeted mutagenesis strategies will promote the study of this mysterious bacterium and facilitate vaccine development against tularemia. Moreover, the general strategy of gene disruption by PCR-based antibiotic resistance cassette insertion is broadly applicable to many bacterial species.

Description
Keywords
Citation
Liu, J., Zogaj, X., Barker, J. R., & Klose, K. E. (2007). Construction of targeted insertion mutations in Francisella tularensis subsp. novicida. BioTechniques, 43(4), 487-492. doi:10.2144/000112574
Department
Molecular Microbiology and Immunology
Neuroscience, Developmental and Regenerative Biology