Abstract
The discovery of the third domain of life, the Archaea, is one of the most exciting findings of the last century. These remarkable prokaryotes are well known for their adaptations to extreme environments; however, Archaea have also conquered moderate environments. Many of the archaeal biochemical processes, such as methane production, are unique in nature and therefore of great scientific interest. Although formerly restricted to biochemical and physiological studies, sophisticated systems for genetic manipulation have been developed during the last two decades for methanogenic archaea, halophilic archaea and thermophilic, sulfur-metabolizing archaea. The availability of these tools has allowed for more complete studies of archaeal physiology and metabolism and most importantly provides the basis for the investigation of gene expression, regulation and function. In this review we provide an overview of methods for genetic manipulation of Methanosarcina spp., a group of methanogenic archaea that are key players in the global carbon cycle and which can be found in a variety of anaerobic environments.
Highlights
Life on earth, as currently perceived by mankind, is encompassed in three major domains Archaea, Eukarya and Bacteria
This review focuses on the genetic manipulation of Methanosarcina spp. and provides an overview about established methods including, random and targeted mutagenesis, complementation, and reporter gene fusions
Recent genetic studies have provided a better understanding of the C1 oxidation/reduction pathway and the energy-conserving electron transport chain of M. bakeri Fusaro (Welander and Metcalf, 2008; Kulkarni et al, 2009)
Summary
As currently perceived by mankind, is encompassed in three major domains Archaea, Eukarya and Bacteria. Archaea share metabolic and physiologic features with Eukarya and Bacteria, but they are unique in many ways (Jarrell et al, 2011; Jun et al, 2011; White, 2011). The study of methanogenic Archaea has a long-standing history because of their central role in the global carbon cycle and their potential application in biofuel production (Fox et al, 1977; Jarrell et al, 2011; De Vrieze et al, 2012). Class III methanogens, Methanosarcinales metabolize a variety of substrates, H2/CO2, C-1 compounds, such as methylamines, methylsulfides or methanol, and acetate in four different methanogenic pathways (Thauer et al, 2008; Anderson et al, 2009). The major difference between these organisms lies within their ability to utilize methanogenic substrates, www.frontiersin.org
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