Candida albicans GFP Strain Construction

Candida albicans wildtype (SC5314) was used for all experiments. For construction of CaGFP (ADH1/adh1::GFP-SAT1) we transformed a cassette including a C. albicans optimized GFP from the vector pNIM1 [55] (link) and SAT1 as selection marker [56] (link) as well as homology regions for integration into the CaADH1 locus into the C. albicans wild type strain SCR5314, using lithium acetate protocol [57] (link). Transformants were grown for two days on YPD with nourseothricine and verified by PCR and microscopy. For an infection of whole blood, C. albicans was grown over night in YPD-medium ( D-glucose, peptone, yeast extract in water) at , reseeded in YPD-medium, grown for five hours at into the mid-log-phase, and harvested in HBSS. C. albicans yeasts were killed by incubation in thimerosal (Sigma-Aldrich) in HBSS at for and then rinsed extensively.

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Hünniger K., Lehnert T., Bieber K., Martin R., Figge M.T, & Kurzai O. (2014). A Virtual Infection Model Quantifies Innate Effector Mechanisms and Candida albicans Immune Escape in Human Blood. PLoS Computational Biology, 10(2), e1003479.

Publication 2014

thimerosal

Blood C albicans Glucose Hbss Infection Lithium acetate Microscopy Peptone Strain Thimerosal Vector Yeasts

Corresponding Organization : Friedrich Schiller University Jena

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Variable analysis

independent variables

Construction of CaGFP (ADH1/adh1::GFP-SAT1)

dependent variables

Transformants were grown for two days on YPD with nourseothricine and verified by PCR and microscopy
Infection of whole blood with C. albicans

control variables

C. albicans wildtype (SC5314) was used for all experiments
C. albicans was grown overnight in YPD-medium (D-glucose, peptone, yeast extract in water) at an unspecified temperature
C. albicans yeasts were killed by incubation in 0.1% thimerosal (Sigma-Aldrich) in HBSS at 37°C for 30 minutes and then rinsed extensively

positive controls

None specified

negative controls

None specified

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