The first transformation method for the thermo-acidophilic archaeon Thermoplasma acidophilum

A transformation method yielding up to 104 transformants per μg circular DNA was developed for Thermoplasma acidophilum. The method is based on a natural DNA uptake process in which T. acidophilum cells keep their integrity and turn competent at pH3.5 and 58°C. Shuttle vector maintenance could not b...

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Bibliographic Details
Published in:Journal of microbiological methods 2013-11, Vol.95 (2), p.145-148
Main Authors: Baka, Erzsébet, Varga, Sándor, Hobel, Cédric, Knispel, Roland Wilhelm, Fekete, Csaba, Ivanics, Milán, Kriszt, Balázs, Nagy, István, Kukolya, József
Format: Article
Language:English
Subjects:
Archaea
Chromosomes, Archaeal - genetics
Cloning, Molecular
Culture Media
DNA, Archaeal - genetics
DNA, Circular - genetics
Drug Resistance, Microbial
Gene Transfer Techniques
Genetic Vectors - genetics
Homologous recombination
Hydrogen-Ion Concentration
Novobiocin
Novobiocin - pharmacology
Plasmid
Promoter Regions, Genetic
Sequence Analysis, DNA
Temperature
Thermoplasma - genetics
Thermoplasma acidophilum
Transformation
Transformation, Genetic
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Summary:A transformation method yielding up to 104 transformants per μg circular DNA was developed for Thermoplasma acidophilum. The method is based on a natural DNA uptake process in which T. acidophilum cells keep their integrity and turn competent at pH3.5 and 58°C. Shuttle vector maintenance could not be detected, since the used NovR gyraseB gene integrated into its chromosomal counterpart by homologous recombination. •A novel transformation method for T. acidophilum was developed.•Culturing the cells on Gelrite-based solid medium was improved.•Novobiocin resistant clones were obtained and selected.•No plasmid maintenance was detected.•Variants of the NovRgyrB gene recombined into the chromosomal gyrB gene.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2013.08.006