Abstract
The tetracycline promoter (tet) has been manipulated in several systems as an inducible regulator of gene expression. In this study the effect of doxycycline on C. glabrata viability has been examined in water and according to CLSI standard method. In addition, flow cytometry analysis was performed to detect SLT2 protein activation by sub-inhibitory concentrations of doxycycline. Our findings show that 100 μg/ml of doxycycline is the maximum tolerated dosage in media; however, C. glabrata was sensitive to 40-50μg/ml in water. 50 μg/ml of doxycycline induced SLT2p phosphorylation and our results support the idea of involvement of doxycycline in alteration of C. glabrata oxidative status. These results should be considered when the tet promoter system is used in yeast molecular studies.
Highlights
Various factors including the increased numbers of immunocompromised patients (e.g. AIDS and cancer patients), a growing aged population, the increased use of indwelling medical devices and antibiotic misuse have led to the increase in Candida infection incidence
Both strains were not sensitive to the highest concentration of doxycycline. Other yeast species such as C. albicans was susceptible to doxycycline at 200 μg/ml [8] when it was tested by using the spotting assay
A study on S. cerevisiae cells treated with 40 μg/ml of doxycycline showed that there was no phenotypic effect and no significant effect on their global genome transcription levels which allow the use of tetO regulatable promoter system for genetic studies [4]
Summary
Various factors including the increased numbers of immunocompromised patients (e.g. AIDS and cancer patients), a growing aged population, the increased use of indwelling medical devices and antibiotic misuse have led to the increase in Candida infection incidence. Detailed genetic studiesof C. glabrata are commonplace thanks to the regulatable promoters which are available to the researchers to induce or repress gene expression in C. glabrata. These promoters, such as tetO, MET3 and PDC1 promoters [2,3] control expression of the gene of interest upon incubation of cells in growth media containing the activating or the repressing substance. The tetregulatable promoteris directed affected by tetracycline,induced by addition of doxycycline to the growth medium with induction being reversed by doxycycline removal. This system has been successfully used forfunctional analysis studies of essential genes with unknown function in C. glabrata [2]. There are no studies that examine the effect of doxycycline on C. glabrata global gene expression
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: International Journal of Clinical & Medical Microbiology
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
