Describe how FISH can be used to determine which cells within a microbial community are actively degrading toluene. Assume the genes involved in the metabolic pathway have been sequenced from several microorganisms and vary only slightly from one another
What will be an ideal response?
Answer: A conserved gene sequence would need to be identified and made into a FISH probe. It is likely CARD-FISH will need to be used to observe a signal. The community could be grown on toluene and exposed to the probe, where microbes expressing the genes for the toluene degradation pathway would hybridize to the CARD-FISH probe and could be visualized using fluorescence microscopy.
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