In vivo transposon tagging in the nonheterocystous nitrogen‐fixing cyanobacterium Leptolyngbya boryana

Nitrogenase is an oxygen‐vulnerable metalloenzyme that catalyzes nitrogen fixation. It largely remains unknown how nitrogenase coexists with oxygenic photosynthesis in nonheterocystous cyanobacteria, since there have been no appropriate model cyanobacteria so far. Here, we demonstrate in vivo transp...

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Bibliographic Details
Published in:FEBS letters 2018-05, Vol.592 (10), p.1634-1642
Main Authors: Tomatsu, Chie, Uesaka, Kazuma, Yamakawa, Hisanori, Tsuchiya, Tohru, Ihara, Kunio, Fujita, Yuichi
Format: Article
Language:English
Subjects:
conjugation
cyanobacteria
genome resequencing
nitrogen fixation
Oxygen Paradox
transposon tagging
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Summary:Nitrogenase is an oxygen‐vulnerable metalloenzyme that catalyzes nitrogen fixation. It largely remains unknown how nitrogenase coexists with oxygenic photosynthesis in nonheterocystous cyanobacteria, since there have been no appropriate model cyanobacteria so far. Here, we demonstrate in vivo transposon tagging in the nonheterocystous cyanobacterium Leptolyngbya boryana as a forward genetics approach. By conjugative transfer, a mini‐Tn5‐derived vector, pKUT‐Tn5‐Sm/Sp, was transferred from Escherichia coli to L. boryana cells. Of 1839 streptomycin‐resistant colonies, we isolated three mutants showing aberrant diazotrophic growth. Genome resequencing identified the insertion sites of the transposon in the mutants. This in vivo transposon tagging mutagenesis of L. boryana provides a promising system to investigate molecular mechanisms to resolve the Oxygen Paradox between nitrogen fixation and oxygenic photosynthesis in cyanobacteria.
ISSN:0014-5793
1873-3468
DOI:10.1002/1873-3468.13079