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In vivo transposon tagging in the nonheterocystous nitrogen‐fixing cyanobacterium Leptolyngbya boryana
Author(s) -
Tomatsu Chie,
Uesaka Kazuma,
Yamakawa Hisanori,
Tsuchiya Tohru,
Ihara Kunio,
Fujita Yuichi
Publication year - 2018
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13079
Subject(s) - transposable element , cyanobacteria , nitrogen fixation , biology , genetics , chemistry , biochemistry , botany , bacteria , gene , genome
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‐Tn 5 ‐derived vector, pKUT ‐Tn 5 ‐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.