Improving the genetic system for Halorubrum lacusprofundi to allow in-frame deletions
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 organis...
| Published in: | Frontiers in Microbiology |
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| Main Authors: | , , |
| Format: | Article in Journal/Newspaper |
| Language: | unknown |
| Published: |
Frontiers Media SA
2023
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| Subjects: | |
| Online Access: | http://dx.doi.org/10.3389/fmicb.2023.1095621 https://www.frontiersin.org/articles/10.3389/fmicb.2023.1095621/full |
| Summary: | 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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