Improving the genetic system for Halorubrum lacusprofundi to allow in-frame deletions.

Gebhard, LJ; Duggin, IG; Erdmann, S

Improving the genetic system for Halorubrum lacusprofundi to allow in-frame deletions.

Gebhard, LJ Duggin, IG Erdmann, S

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Publisher:: FRONTIERS MEDIA SA
Publication Type:: Journal Article
Citation:: Front Microbiol, 2023, 14, pp. 1095621
Issue Date:: 2023

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Field	Value	Language
dc.contributor.author	Gebhard, LJ
dc.contributor.author	Duggin, IG
dc.contributor.author	Erdmann, S
dc.date.accessioned	2024-01-11T03:49:21Z
dc.date.available	2023-03-13
dc.date.available	2024-01-11T03:49:21Z
dc.date.issued	2023
dc.identifier.citation	Front Microbiol, 2023, 14, pp. 1095621
dc.identifier.issn	1664-302X
dc.identifier.issn	1664-302X
dc.identifier.uri	http://hdl.handle.net/10453/174300
dc.description.abstract	Halorubrum lacusprofundi is a cold-adapted halophilic archaeon isolated from Deep Lake, Antarctica. Hrr. lacusprofundi is commonly used to study adaptation to cold environments and thereby a potential source for biotechnological products. Additionally, in contrast to other haloarchaeal model organisms, Hrr. lacusprofundi is also susceptible to a range of different viruses and virus-like elements, making it a great model to study virus-host interactions in a cold-adapted organism. A genetic system has previously been reported for Hrr. lacusprofundi; however, it does not allow in-frame deletions and multiple gene knockouts. Here, we report the successful generation of uracil auxotrophic (pyrE2) mutants of two strains of Hrr. lacusprofundi. Subsequently, we attempted to generate knockout mutants using the auxotrophic marker for selection. However, surprisingly, only the combination of the auxotrophic marker and antibiotic selection allowed the timely and clean in-frame deletion of a target gene. Finally, we show that vectors established for the model organism Haloferax volcanii are deployable for genetic manipulation of Hrr. lacusprofundi, allowing the use of the portfolio of genetic tools available for H. volcanii in Hrr. lacusprofundi.
dc.format	Electronic-eCollection
dc.language	eng
dc.publisher	FRONTIERS MEDIA SA
dc.relation	http://purl.org/au-research/grants/arc/FT160100010
dc.relation.ispartof	Front Microbiol
dc.relation.isbasedon	10.3389/fmicb.2023.1095621
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0502 Environmental Science and Management, 0503 Soil Sciences, 0605 Microbiology
dc.subject.classification	3107 Microbiology
dc.subject.classification	3207 Medical microbiology
dc.title	Improving the genetic system for Halorubrum lacusprofundi to allow in-frame deletions.
dc.type	Journal Article
utslib.citation.volume	14
utslib.location.activity	Switzerland
utslib.for	0502 Environmental Science and Management
utslib.for	0503 Soil Sciences
utslib.for	0605 Microbiology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - AIMI - Australian Institute for Microbiology & Infection
utslib.copyright.status	open_access	*
dc.date.updated	2024-01-11T03:49:14Z
pubs.publication-status	Published online
pubs.volume	14

Abstract:

Halorubrum lacusprofundi is a cold-adapted halophilic archaeon isolated from Deep Lake, Antarctica. Hrr. lacusprofundi is commonly used to study adaptation to cold environments and thereby a potential source for biotechnological products. Additionally, in contrast to other haloarchaeal model organisms, Hrr. lacusprofundi is also susceptible to a range of different viruses and virus-like elements, making it a great model to study virus-host interactions in a cold-adapted organism. A genetic system has previously been reported for Hrr. lacusprofundi; however, it does not allow in-frame deletions and multiple gene knockouts. Here, we report the successful generation of uracil auxotrophic (pyrE2) mutants of two strains of Hrr. lacusprofundi. Subsequently, we attempted to generate knockout mutants using the auxotrophic marker for selection. However, surprisingly, only the combination of the auxotrophic marker and antibiotic selection allowed the timely and clean in-frame deletion of a target gene. Finally, we show that vectors established for the model organism Haloferax volcanii are deployable for genetic manipulation of Hrr. lacusprofundi, allowing the use of the portfolio of genetic tools available for H. volcanii in Hrr. lacusprofundi.

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http://hdl.handle.net/10453/174300