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Study of Fe-S Cluster Proteins in Methanococcus maripaludis , a Model Archaeal Organism
Cuiping Zhao1, Christina A Roberts2, Ian J Drake3
1Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, NJ, USA.
Abstract:
Iron-sulfur (Fe-S) clusters are among the oldest and most versatile cofactors present in all domains of life. Many bacterial and eukaryotic Fe-S proteins have been well-characterized, whereas the archaeal ones are less studied. Fe-S proteins are particularly abundant and play essential roles in methanogenic archaea. Methanococcus maripaludis is a model methanogen with available genetic tools. Here, we describe the techniques for anaerobic cultivation of M. maripaludis with formate, liposome-mediated transformation, expression and anoxic affinity purification of Fe-S proteins, Fe-S cluster reconstitution, and analysis of Fe-S proteins by UV-visible absorption spectroscopy.
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