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.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Liu J,Zogaj X,Barker JR,Klose KEdoi
10.2144/000112574subject
Has Abstractpub_date
2007-10-01 00:00:00pages
487-90, 492issue
4eissn
0736-6205issn
1940-9818pii
000112574journal_volume
43pub_type
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