TY - CHAP
T1 - Genetic methods for Methanosarcina species
AU - Buan, Nicole
AU - Kulkarni, Gargi
AU - Metcalf, William
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
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U2 - 10.1016/B978-0-12-385112-3.00002-0
DO - 10.1016/B978-0-12-385112-3.00002-0
M3 - Chapter
C2 - 21402208
AN - SCOPUS:79952613800
T3 - Methods in Enzymology
SP - 23
EP - 42
BT - Methods in Enzymology
PB - Academic Press Inc.
ER -