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Supplemental Information

Table 1.Primers used throughout this study

DOI: 10.7287/peerj.preprints.177v1/supp-1

Figure 1. Regulation of M. smegmatis adenylate cyclase genes under starvation conditions.

Semi-quantitative RT-PCR was used to compare adenylate cyclase mRNA levels between late-log phase cultures incubated for 4 hours in mycomedia (control) or PBS (starvation). (A) A representative depiction of PCR amplified cDNA separated using agarose gel electrophoresis. (B) Data obtained from ImageJ quantifying the band densities observed on the depicted agarose gel comparing the cDNA levels under control (black bars) and starvation (white bars) conditions. (C) Average of three different experiments represented as fold differences of starvation compared to control expression. * indicates statistically significant difference between expression in control and starvation for an individual gene (p<0.05)

DOI: 10.7287/peerj.preprints.177v1/supp-2

Table 2. Percent identity between Mtb and M. smegmatis ortholog promoters

DOI: 10.7287/peerj.preprints.177v1/supp-3

Figure 2. Regulation of M. smegmatis adenylate cyclase genes under low pH and oxidative stress

Semi-quantitative RT-PCR was used to compare adenylate cyclase mRNA levels between late-log phase cultures incubated for 4 hours in mycomedia (control, black bars) , mycomedia adjusted to pH 5.5 (low pH, hatched bars), and mycomedia containing H2O2 (H2O2, grey bars). Results are the average of three independent experiments and are expressed in fold difference comparing experimental condition to control. * indicates conditions with statistically significant differences in expression compared to control for an individual gene (p<0.05)

DOI: 10.7287/peerj.preprints.177v1/supp-4

Figure 3. Regulation of M. tuberculosis adenylate cyclase genes.

GFP: promoter fusions containing M. tuberculosis promoters were electroporated into M. smegmatis and used to examine adenylate cyclase expression in late log phase cultures after 4 hour incubation in mycomedia (control, black bar), PBS (starvation, white bar), mycomedia adjusted to pH 5.5 (low pH, hatched bars), and mycomedia containing H2O2 (H2O2, grey bars). Fluorescence was normalized to 106 cells and represented as the average of three independent experiments. * indicates conditions with statistically significant differences in expression compared to control for an individual gene (p<0.05)

DOI: 10.7287/peerj.preprints.177v1/supp-5

Additional Information

Competing Interests

The authors have no competing interests that would bias this work.

Author Contributions

Sarah J Casey performed the experiments, analyzed the data, wrote the paper.

Mica J Ford performed the experiments.

Michaela Gazdik conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper.

Grant Disclosures

The following grant information was disclosed by the authors:

National Institute of Allergy and Infectious Diseases grant number 1R15AI084058-01

Funding

This work was supported by National Institutes of Health Grant 1R15AI084058-01. Additional support was obtained through internal Ferrum College research grants. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.


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