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 an...

background: causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. brucellosis is the most common zoonotic infectious disease that would cause great economic losses. thus, recognition of pathogenic and immunogenic factors in the genus brucella can lead to control this health problem. objectives: considering the importanc...

A rapid method is described to efficiently perform site-directed mutagenesis based on overlap extension polymerase chain reaction (OE-PCR). Two template DNA molecules in different orientations relative to only one universal primer were amplified in parallel. By choosing a high dilution of mutagenic primers it was possible to run an overlap extension PCR in only one reaction without purification...