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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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Published in: | FEBS letters 2018-05, Vol.592 (10), p.1634-1642 |
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creator | Tomatsu, Chie Uesaka, Kazuma Yamakawa, Hisanori Tsuchiya, Tohru Ihara, Kunio Fujita, Yuichi |
description | 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. |
doi_str_mv | 10.1002/1873-3468.13079 |
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subjects | conjugation cyanobacteria genome resequencing nitrogen fixation Oxygen Paradox transposon tagging |
title | In vivo transposon tagging in the nonheterocystous nitrogen‐fixing cyanobacterium Leptolyngbya boryana |
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