Molecular toolbox for genetic manipulation of the stalked budding bacterium Hyphomonas neptunium.
Appl Environ Microbiol
; 81(2): 736-44, 2015 Jan.
Article
in En
| MEDLINE
| ID: mdl-25398860
ABSTRACT
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Alphaproteobacteria
/
Genetics, Microbial
/
Molecular Biology
Language:
En
Journal:
Appl Environ Microbiol
Year:
2015
Document type:
Article
Affiliation country: