Molecular toolbox for genetic manipulation of the stalked budding bacterium Hyphomonas neptunium.
The alphaproteobacterium Hyphomonas neptunium proliferates by a unique budding mechanism in which daughter cells emerge from the end of a stalk-like extension emanating from the mother cell body. Studies of this species so far have been hampered by the lack of a genetic system and of molecular tools allowing the regulated expression of target genes. Based on microarray analyses, this work identifies two H. neptunium promoters that are activated specifically by copper and zinc. Functional analyses show that they have low basal activity and a high dynamic range, meeting the requirements for use as a multipurpose expression system. To facilitate their application, the two promoters were incorporated into a set of integrative plasmids, featuring a choice of two different selection markers and various fluorescent protein genes. These constructs enable the straightforward generation and heavy metal-inducible synthesis of fluorescent protein fusions in H. neptunium, thereby opening the door to an in-depth analysis of polar growth and development in this species.
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Alexandra
A
Jung
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Sabrina
S
Eisheuer
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Emöke
E
Cserti
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Oliver
O
Leicht
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Wolfgang
W
Strobel
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Andrea
A
Möll
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Susan
S
Schlimpert
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany.
Juliane
J
Kühn
Max Planck Institute for Terrestrial Microbiology, Marburg, Germany Faculty of Biology, Philipps-Universität, Marburg, Germany LOEWE Center for Synthetic Microbiology, Marburg, Germany thanbichler@uni-marburg.de.
Martin
M
Thanbichler
D020561:Alphaproteobacteria / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008670:Metals / Q000378:metabolism; D046228:Microarray Analysis; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA; D015533:Transcriptional Activation / Q000187:drug effects
D004269:DNA, Bacterial; D008164:Luminescent Proteins; D008670:Metals