{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T02:32:16Z","timestamp":1791340336289,"version":"4.3.1"},"reference-count":41,"publisher":"Microbiology Society","issue":"4","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2009,4,1]]},"abstract":"<jats:p>\n                  <jats:italic>Francisella tularensis<\/jats:italic> is a fastidious Gram-negative bacterium responsible for the zoonotic disease tularemia. Investigation of the biology and molecular pathogenesis of <jats:italic>F. tularensis<\/jats:italic> has been limited by the difficulties in manipulating such a highly pathogenic organism and by a lack of genetic tools. However, recent advances have substantially improved the ability of researchers to genetically manipulate this organism. To expand the molecular toolbox we have developed two systems to stably integrate genetic elements in single-copy into the <jats:italic>F. tularensis<\/jats:italic> genome. The first system is based upon the ability of transposon Tn<jats:italic>7<\/jats:italic> to insert in both a site- and orientation-specific manner at high frequency into the <jats:italic>att<\/jats:italic>Tn<jats:italic>7<\/jats:italic> site located downstream of the highly conserved <jats:italic>glmS<\/jats:italic> gene. The second system consists of a <jats:italic>sacB<\/jats:italic>-based suicide plasmid used for allelic exchange of unmarked elements with the <jats:italic>blaB<\/jats:italic> gene, encoding a <jats:italic>\u03b2<\/jats:italic>-lactamase, resulting in the replacement of <jats:italic>blaB<\/jats:italic> with the element and the loss of ampicillin resistance. To test these new tools we used them to complement a novel <jats:sc>d<\/jats:sc>-glutamate auxotroph of <jats:italic>F. tularensis<\/jats:italic> LVS, created using an improved <jats:italic>sacB<\/jats:italic>-based allelic exchange plasmid. These new systems will be helpful for the genetic manipulation of <jats:italic>F. tularensis<\/jats:italic> in studies of tularemia biology, especially where the use of multi-copy plasmids or antibiotic markers may not be suitable.<\/jats:p>","DOI":"10.1099\/mic.0.022491-0","type":"journal-article","created":{"date-parts":[[2009,3,31]],"date-time":"2009-03-31T07:52:51Z","timestamp":1238485971000},"page":"1152-1163","source":"Crossref","is-referenced-by-count":68,"title":["Single-copy chromosomal integration systems for Francisella tularensis"],"prefix":"10.1099","volume":"155","author":[{"given":"Eric D.","family":"LoVullo","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia R.","family":"Molins-Schneekloth","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology, and Pathology, 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