The genetic toolbox for Acidovorax temperans

Environmental isolates belonging to the genus Acidovorax play a crucial role in degrading a wide range of pollutants. Studies on Acidovorax are currently limited for many species due to the lack of genetic tools. Here, we described the use of the replicon from a small, cryptic plasmid indigenous to...

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Bibliographic Details
Published in:Journal of microbiological methods 2015-08, Vol.115, p.129-138
Main Authors: Boycheva, S.S., Pichler, F.B., Heijstra, B.D., Lau, K.E.M., Turner, S.J.
Format: Article
Language:English
Subjects:
Acidovorax
Comamonadaceae - genetics
Comamonadaceae - metabolism
Counterselectable marker
Genetic Techniques
Genetic Vectors - genetics
Genetic Vectors - metabolism
gfp
Indigenous plasmid
Plasmids - genetics
Plasmids - metabolism
Recombination
Replicon
Scarless
Shuttle vector
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Summary:Environmental isolates belonging to the genus Acidovorax play a crucial role in degrading a wide range of pollutants. Studies on Acidovorax are currently limited for many species due to the lack of genetic tools. Here, we described the use of the replicon from a small, cryptic plasmid indigenous to Acidovorx temperans strain CB2, to generate stably maintained shuttle vectors. In addition, we have developed a scarless gene knockout technique, as well as establishing green fluorescent protein (GFP) reporter and complementation systems. Taken collectively, these tools will improve genetic manipulations in the genus Acidovorax. •We found a small, cryptic plasmid indigenous to Acidovorax temperans CB2 strain.•A set of shuttle vectors was constructed.•We developed a scarless gene knockout technique based on homologous recombination.•A gfp reporter system was established.•The reported tools will improve genetic studies in the genus Acidovorax.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2015.06.006