PNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans

Mitchell T. Shea, Mary E. Walter, Nikolas Duszenko, Anne Lise Ducluzeau, Jared Aldridge, Shannon K. King, Nicole R. Buan

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available Escherichia coli plasmid, pNEB193, and cannot replicate autonomously in methanogens. The designed plasmids facilitate markerless gene deletion, gene transcription, protein expression, and purification of proteins with cleavable affinity tags from the methanogen, M. acetivorans.

Original languageEnglish (US)
Pages (from-to)27-35
Number of pages9
JournalPlasmid
Volume84-85
DOIs
StatePublished - Mar 1 2016

Fingerprint

Methanosarcina
Archaea
Methane
Gene Deletion
Suicide
Plasmids
Proteins
Coenzymes
Post Translational Protein Processing
Escherichia coli
Bacteria
Gene Expression
Enzymes

Keywords

  • Archaea
  • Methanogen
  • Methanosarcina
  • Protein expression

ASJC Scopus subject areas

  • Molecular Biology

Cite this

PNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans. / Shea, Mitchell T.; Walter, Mary E.; Duszenko, Nikolas; Ducluzeau, Anne Lise; Aldridge, Jared; King, Shannon K.; Buan, Nicole R.

In: Plasmid, Vol. 84-85, 01.03.2016, p. 27-35.

Research output: Contribution to journalArticle

Shea, Mitchell T. ; Walter, Mary E. ; Duszenko, Nikolas ; Ducluzeau, Anne Lise ; Aldridge, Jared ; King, Shannon K. ; Buan, Nicole R. / PNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans. In: Plasmid. 2016 ; Vol. 84-85. pp. 27-35.
@article{d67a3979b9b246adb6e8e20e87a54ea8,
title = "PNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans",
abstract = "Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available Escherichia coli plasmid, pNEB193, and cannot replicate autonomously in methanogens. The designed plasmids facilitate markerless gene deletion, gene transcription, protein expression, and purification of proteins with cleavable affinity tags from the methanogen, M. acetivorans.",
keywords = "Archaea, Methanogen, Methanosarcina, Protein expression",
author = "Shea, {Mitchell T.} and Walter, {Mary E.} and Nikolas Duszenko and Ducluzeau, {Anne Lise} and Jared Aldridge and King, {Shannon K.} and Buan, {Nicole R.}",
year = "2016",
month = "3",
day = "1",
doi = "10.1016/j.plasmid.2016.02.003",
language = "English (US)",
volume = "84-85",
pages = "27--35",
journal = "Plasmid",
issn = "0147-619X",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - PNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans

AU - Shea, Mitchell T.

AU - Walter, Mary E.

AU - Duszenko, Nikolas

AU - Ducluzeau, Anne Lise

AU - Aldridge, Jared

AU - King, Shannon K.

AU - Buan, Nicole R.

PY - 2016/3/1

Y1 - 2016/3/1

N2 - Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available Escherichia coli plasmid, pNEB193, and cannot replicate autonomously in methanogens. The designed plasmids facilitate markerless gene deletion, gene transcription, protein expression, and purification of proteins with cleavable affinity tags from the methanogen, M. acetivorans.

AB - Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available Escherichia coli plasmid, pNEB193, and cannot replicate autonomously in methanogens. The designed plasmids facilitate markerless gene deletion, gene transcription, protein expression, and purification of proteins with cleavable affinity tags from the methanogen, M. acetivorans.

KW - Archaea

KW - Methanogen

KW - Methanosarcina

KW - Protein expression

UR - http://www.scopus.com/inward/record.url?scp=84958559359&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84958559359&partnerID=8YFLogxK

U2 - 10.1016/j.plasmid.2016.02.003

DO - 10.1016/j.plasmid.2016.02.003

M3 - Article

C2 - 26876941

AN - SCOPUS:84958559359

VL - 84-85

SP - 27

EP - 35

JO - Plasmid

JF - Plasmid

SN - 0147-619X

ER -