Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix

Christiansen G, Goesmann A, Kurmayer R (2014)
Applied and environmental microbiology 80(16): 4887-4897.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Several gene clusters that are responsible for toxin synthesis in bloom-forming cyanobacteria have been found to be associated with transposable elements (TEs). In particular, insertion sequence (IS) elements were shown to play a role in the inactivation or recombination of the genes responsible for cyanotoxin synthesis. Plasmids have been considered important vectors of IS element distribution to the host. In this study, we aimed to elucidate the IS elements propagated on the plasmids and the chromosome of the toxic cyanobacterium Planktothrix agardhii NIVA-CYA126/8 by means of high-throughput sequencing. In total, five plasmids (pPA5.5, pPA14, pPA50, pPA79, and pPA115, of 5, 6, 50, 79, and 120 kbp, respectively) were elucidated, and two plasmids (pPA5.5, pPA115) were found to propagate full IS element copies. Large stretches of shared DNA information between plasmids were constituted of TEs. Two plasmids (pPA5.5, pPA14) were used as candidates to engineer shuttle vectors (named pPA5.5SV and pPA14SV, respectively) in vitro by PCR amplification and the subsequent transposition of the Tn5 cat transposon containing the R6Kγ origin of replication of Escherichia coli. While pPA5.5SV was found to be fully segregated, pPA14SV consistently co-occurred with its wild-type plasmid even under the highest selective pressure. Interestingly, the Tn5 cat transposon became transferred by homologous recombination into another plasmid, pPA50. The availability of shuttle vectors is considered to be of relevance in investigating genome plasticity as a consequence of homologous recombination events. Combining the potential of high-throughput sequencing and in vitro production of shuttle vectors makes it simple to produce species-specific shuttle vectors for many cultivable prokaryotes.
Erscheinungsjahr
2014
Zeitschriftentitel
Applied and environmental microbiology
Band
80
Ausgabe
16
Seite(n)
4887-4897
ISSN
0099-2240
eISSN
1098-5336
Page URI
https://pub.uni-bielefeld.de/record/2690174

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Christiansen G, Goesmann A, Kurmayer R. Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix. Applied and environmental microbiology. 2014;80(16):4887-4897.
Christiansen, G., Goesmann, A., & Kurmayer, R. (2014). Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix. Applied and environmental microbiology, 80(16), 4887-4897. doi:10.1128/AEM.01188-14
Christiansen, G., Goesmann, A., and Kurmayer, R. (2014). Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix. Applied and environmental microbiology 80, 4887-4897.
Christiansen, G., Goesmann, A., & Kurmayer, R., 2014. Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix. Applied and environmental microbiology, 80(16), p 4887-4897.
G. Christiansen, A. Goesmann, and R. Kurmayer, “Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix”, Applied and environmental microbiology, vol. 80, 2014, pp. 4887-4897.
Christiansen, G., Goesmann, A., Kurmayer, R.: Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix. Applied and environmental microbiology. 80, 4887-4897 (2014).
Christiansen, Guntram, Goesmann, Alexander, and Kurmayer, Rainer. “Elucidation of insertion elements carried on plasmids and in vitro construction of shuttle vectors from the toxic cyanobacterium planktothrix”. Applied and environmental microbiology 80.16 (2014): 4887-4897.

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