Open AccessImproved shuttle vectors for Francisella tularensis genetics
Author(s) -
LoVullo Eric D.,
Sherrill Lani A.,
Pavelka Martin S.
Publication year - 2009
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2008.01440.x
Subject(s) - francisella tularensis , shuttle vector , francisella , plasmid , multiple cloning site , biology , genetics , cloning (programming) , vector (molecular biology) , tularemia , restriction enzyme , escherichia coli , gene , recombinant dna , microbiology and biotechnology , virulence , computer science , programming language
We previously described the construction and characterization of Escherichia coli–Francisella tularensis shuttle vectors, derived from the cryptic Francisella plasmid pFNL10, for the genetic manipulation of F. tularensis ssp. tularensis . We now report further characterization of the biology of these shuttle vectors and the development of a new generation of Francisella plasmids. We show that the addition of ORF3 from pFNL10 can convert an unstable shuttle vector into a stable one, and that this is likely due to increased plasmid copy number. We also describe various improvements to the earlier generations of shuttle vectors, such as the addition of a multiple cloning site containing a novel RsrII restriction endonuclease site for directional insertion of Francisella genes, and the inclusion of the F. tularensis blaB promoter for heterologous gene expression.