The Development of Fluorescence-based Molecular Tools for Studying Extremophiles

Lead Investigator: Don Benn, University of Montana

Studies of the Earth�s extreme environments and the organisms that live within these �harsh� habitats provide opportunities to gain insight into the biodiversity on earth and the types of life forms that may exist on other planets.  Earth�s hot volcanic beds, where extremophiles like the Sulfolobus species (and the viruses that infect them) exist, are an example of one type of extreme environment where thermophilic life survives and thrive.  Understanding the biomolecular and genetic substrates that allow microbial life to exist under extreme conditions, requires molecular tools.  Due to the unique characteristics of hyperthermophilic archaea, such as Sulfolobus, standard molecular tools and techniques, used in the study mesophilic bacteria, are inadequate.  In fact, there are very few genetic tools available to study hyperthermophilic archaea.  A System that employs transformable plasmids endowed with an inducible green fluorescent protein (GFP) gene and selection marker that can withstand high temperatures and acidic environments would serve as very useful tool for studying these thermophiles.  A super--‐folded GFP (sGFP) has been recently tested to 75�C, Which is in the lower physiological range of Sulfolobus.  The Goal of this MSGC Research initiation proposal is to develop a sGFP-�based molecular tool for monitoring cellular and biomolecular activity in Sulfolobus And to test this construct�s utility in monitoring hyperthermophilic virus-�host interactions and in addressing hypotheses related to hyperthermophile protein function and cellular localization.  This Research is in line with Goal 5 Of the NASA Astrobiology Institute�s Astrobiology Roadmap.  Given That Yellowstone National Park (Montana/Wyoming)  Is a large geothermal area that features Sulfolobus habitats, this research is also relevant to sciences in the state of  Montana.