Single molecule FISH for the quantitative detection of mRNA transcripts targeting genes relevant for mercury methylation in Desulfovibrio desulfuricans ND132
EMSL Project ID
50065
Abstract
Mercury (Hg) methylation by anaerobic bacteria and archaea is the primary source of toxic methylmercury (MeHg), which bioaccumulates up trophic levels and affects humans through consumption of fish and other seafood. A two-gene cluster, consisting of hgcA and hgcB, encodes two of the proteins essential for this mercury methylation activity in these microorganisms. However, our understanding of factors controlling the expression of genes required for mercury methylation is currently limited. We hypothesize that mercury methylation activity is controlled on the level of transcription. Specific aim: We propose to use quantitative fluorescence in situ hybridization (FISH) in combination with stochastic optical reconstruction microscopy (STORM) imaging to determine transcript levels for genes of interest in single cells. This proof-of-principle experiment will demonstrate the feasibility of FISH-STORM for accurately quantifying transcript levels in single cells of the model methylator Desulfovibrio desulfuricans ND132. Probing transcript levels and factors regulating gene expression in methylating bacteria is expected to enable a more accurate determination of the mercury methylation potential for specific microbial species in environmental samples.
Project Details
Project type
Limited Scope
Start Date
2018-05-29
End Date
2018-07-29
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
Team Members