Abstract
Here we present the characterization of a Francisella bacteriophage (vB_FhiM_KIRK) including the morphology, the genome sequence and the induction of the prophage. The prophage sequence (FhaGI-1) has previously been identified in F. hispaniensis strain 3523. UV radiation induced the prophage to assemble phage particles consisting of an icosahedral head (~52 nm in diameter), a tail of up to 97 nm in length and a mean width of 9 nm. The double stranded genome of vB_FhiM_KIRK contains 51 open reading frames and is 34,259 bp in length. The genotypic and phylogenetic analysis indicated that this phage seems to belong to the Myoviridae family of bacteriophages. Under the conditions tested here, host cell (Francisella hispaniensis 3523) lysis activity of KIRK was very low, and the phage particles seem to be defective for infecting new bacterial cells. Nevertheless, recombinant KIRK DNA was able to integrate site-specifically into the genome of different Francisella species after DNA transformation.
Highlights
Francisella tularensis is a Gram-negative zoonotic bacterium able to cause tularemia in a wide range of animals and in humans, where it causes various clinical expressions ranging from skin lesions to severe pneumonia, depending on the route of infection [1,2]
E. coli strains were grown at 37 ◦C in lysogeny broth (LB; 1% bacto tryptone, 0.5% yeast extract, 0.5% NaCl) or on LB agar (LB supplemented with 1.2% agar)
The putative prophage DNA sequence starts with the attL site, which is a part of the bacterial tRNA-Val gene and is located upstream of protein Fhv_0001 (= FN3523_0986 of the prophage)
Summary
Francisella tularensis is a Gram-negative zoonotic bacterium able to cause tularemia in a wide range of animals and in humans, where it causes various clinical expressions ranging from skin lesions to severe pneumonia, depending on the route of infection [1,2]. Infections in humans are mostly associated with the highly virulent F. tularensis subsp. Fhi strain 3523 was isolated from a patient infected by this bacterium as a result of a cut received in brackish water in North Territory of Australia and was initially believed to be a Fno-like species [6]. It was the first reported Francisella strain from the Southern hemisphere. The putative prophage is not present in the genome sequence of the Fhi strain FSC454
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