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Table 1 Genetic elements used for manipulation of methanogens

From: Methanogens: biochemical background and biotechnological applications

Element(s)

Type

Use/phenotype

Organisma

References

(Counter-) selectable markers

 pac

Resistance gene

Puromycin resistance

Methanococcus, Methanosarcina

Gernhardt et al. (1990) and Metcalf et al. (1997)

 APH3’II

Resistance gene

Neomycin resistance

M. maripaludis

Argyle et al. (1996)

 apH-2b

Resistance gene

Neomycin resistance

M. mazei b

Mondorf et al. (2012)

 ileS3

Resistance gene

Pseudomonic acid resistance

M. acetivorans

Boccazzi et al. (2000)

 serC

Biosynthesis gene

Auxotrophic marker

M. barkeri

Metcalf et al. (1996)

 proC

Biosynthesis gene

Auxotrophic marker

M. acetivorans

Pritchett et al. (2004)

 hpt

Purine salvage gene

Counter-selectable marker (8-aza-2,6-diaminopurine)

Methanosarcina

Pritchett et al. (2004)

 upt

Purine salvage gene

Counter-selectable marker (6-aza-uracil)

M. maripaludis

Moore and Leigh (2005)

Controlling gene expression

 PhmvA

Promoter

Constitutive gene expression

Methanococcus

Beneke et al. (1995)

 Psl

Promoter

Constitutive gene expression

Methanococcus

Sun and Klein (2004)

 PmcrB/Tmcr

Promoter/terminator

Constitutive gene expression

Methanococcus, Methanosarcina

Gernhardt et al. (1990) and MÅ«ller et al. (1985)

 PmcrB(tetO)/TetR

Promoter/repressor

Tetracycline-dependent gene de-repression

Methanosarcina

Guss et al. (2008)

 PmtaCB1

Promoter

Methanol-dependent gene induction

M. acetivorans

Rother et al. (2005)

 PmtbC

Promoter

Methylamine-dependent gene induction

M. mazei

Mondorf et al. (2012)

 Pnif/NrpR

Promoter/repressor

N-dependent gene repression

M. maripaludis

Cohen-Kupiec et al. (1997 and Lie and Leigh (2003)

 tc-RS4

Riboswitch

Tetracycline-dependent gene repression

M. acetivorans

Demolli et al. (2014)

Random mutagenesis

 Himar1/Tnp

Insect transposon/transposase

Random transposon mutagenesis

M. acetivorans, M. maripaludis

Zhang et al. (2000) and Sattler et al. (2013)

 Tn5/Tnp

Transposon/transposase

Random transposon mutagenesis

M. maripaludis

Porat and Whitman (2009)

Reporter genes

 lacZ

Reporter gene

Quantification of gene expression/promoter function

M. maripaludis

Gardner and Whitman (1999)

 uidA

Reporter gene

Quantification of gene expression/promoter function

M. voltae, Methanosarcina

Beneke et al. (1995) and Pritchett et al. (2004)

 bla

Reporter gene

Quantification of gene expression/promoter function

M. acetivorans

Demolli et al. (2014)

Facilitating recombination/mutagenesis

 \(\Phi {\text{C}}31\)

 int

 attB/P

Phage recombination system

Site specific chromosomal integration

Methanosarcina

Guss et al. (2008)

 flp/FRT

Yeast recombination system

Marker rescue

Methanosarcina, Methanococcus

Welander and Metcalf (2008) and Hohn et al. (2011)

 Cas9

Nuclease

Targeted genome editing

M. acetivorans

Nayak and Metcalf (2017)

  1. aIf genus is given, the element is effective in various model species
  2. bThe construct used does not confer resistance to neomycin in M. acetivorans (Matschiavelli and Rother, unpublished)