Genetic methods for Methanosarcina species

Unlike most methanogenic microorganisms, Methanosarcina species are capable of utilizing a variety of growth substrates, a trait that greatly simplifies genetic analysis of the methanogenic process. The genetic tools and techniques discussed in this chapter form the basis for all genetic experiments in Methanosarcina acetivorans C2A and Methanosarcina barkeri Fusaro, two methanogens that are routinely used as model organisms for genetic experiments. Based on a number of reports, it is likely that they are portable to other Methanosarcina species, and perhaps to other methanogens as well. Here, we outline the procedures for high-efficiency transformation using liposomes, gene expression from a plasmid, and exploitation of homologous and site-specific recombination to add and delete genes from the chromosome. Finally, we outline the method for testing whether a gene is essential. These methods can be adapted and combined in any number of ways to design genetic experiments in Methanosarcina.