Cultivation of Francisella tularensis Strains

The Francisella tularensis subsp. holarctica live vaccine strain (LVS) was obtained from the Centers for Disease Control and Prevention (Atlanta, GA), and F. tularensis subsp. tularensis Schu S4 was obtained from BEI Resources. Wild-type (WT) F. tularensis Schu S4 was initially cultured on chocolate agar supplemented with 1% IsoVitaleX (see the supplemental material for the recipe). WT F. tularensis Schu S4 harboring a luciferase (LUX) plasmid (pJB3 [29 (link)]) and WT F. tularensis LVS pEDL20 (53 (link)) were cultured on chocolate agar supplemented with 1% IsoVitaleX and 200 μg/mL hygromycin (Cayman Chemicals). WT F. tularensis Schu S4 expressing green fluorescent protein (GFP) was cultured on chocolate agar supplemented with 1% IsoVitaleX and 10 μg/mL kanamycin (Sigma-Aldrich). All strains were grown overnight at 37°C with aeration in Chamberlain’s defined broth medium (CDM) following growth on plates (54 (link)).

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Dominguez S.R., Whiles S., Deobald K.N, & Kawula T. (2022). Francisella tularensis Exploits AMPK Activation to Harvest Host-Derived Nutrients Liberated from Host Lipolysis. Infection and Immunity, 90(8), e00155-22.

Publication 2022

hygromycin kanamycin

Agar Cayman Chocolate Francisella tularensis subsp holarctica Green fluorescent protein Growth plates Hygromycin Kanamycin Live vaccine Luciferase Plasmid Strains

Corresponding Organization : Washington State University

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Variable analysis

independent variables

Strain of Francisella tularensis used (F. tularensis subsp. holarctica live vaccine strain (LVS), F. tularensis subsp. tularensis Schu S4, and F. tularensis Schu S4 expressing luciferase or green fluorescent protein)
Culture media (chocolate agar supplemented with 1% IsoVitaleX, with or without antibiotics)

dependent variables

Not explicitly mentioned

control variables

Growth conditions (overnight at 37°C with aeration in Chamberlain's defined broth medium (CDM) following growth on plates)
Use of positive and negative controls

controls

Positive control: Wild-type (WT) F. tularensis Schu S4
Negative control: Not specified

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