Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors

Gren T, Ortseifen V, Wibberg D, Schneiker-Bekel S, Bednarz H, Niehaus K, Zemke T, Persicke M, Pühler A, Kalinowski J (2016)
Journal of Biotechnology 232: 79-88.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
The alpha-glucosidase inhibitor acarbose is used for treatment of diabetes mellitus type II, and is manufactured industrially with overproducing derivatives of Actinoplanes sp. SE50/110, reportedly obtained by conventional mutagenesis. Despite of high industrial significance, only limited information exists regarding acarbose metabolism, function and regulation of these processes, due to the absence of proper genetic engineering methods and tools developed for this strain. Here, a basic toolkit for genetic engineering of Actinoplanes sp. SE50/110 was developed, comprising a standardized protocol for a DNA transfer through Escherichia coli-Actinoplanes intergeneric conjugation and applied for the transfer of phi C31, phi BT1 and VWB actinophage-based integrative vectors. Integration sites, occurring once per genome for all vectors, were sequenced and characterized for the first time in Actinoplanes sp. SE50/110. Notably, in case of phi C31 based vector pSET152, the integration site is highly conserved, while for phi BT1 and the VWB based vectors pRT801 and pSOK804, respectively, no sequence similarities to those in other bacteria were detected. The studied plasmids were proven to be stable and neutral with respect to strain morphology and acarbose production, enabling future use for genetic manipulations of Actinoplanes sp. SE50/110. To further broaden the spectrum of available tools, a GUS reporter system, based on the pSET152 derived vector, was also established in Actinoplanes sp. SE50/110. (C) 2016 Elsevier B.V. All rights reserved.
Erscheinungsjahr
Zeitschriftentitel
Journal of Biotechnology
Band
232
Seite
79-88
Konferenz
10th CeBiTec Symposium on Bioinformatics for Biotechnology and Biomedicine
Konferenzort
Bielefeld Univ, Bielefeld, GERMANY
ISSN
eISSN
PUB-ID

Zitieren

Gren T, Ortseifen V, Wibberg D, et al. Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors. Journal of Biotechnology. 2016;232:79-88.
Gren, T., Ortseifen, V., Wibberg, D., Schneiker-Bekel, S., Bednarz, H., Niehaus, K., Zemke, T., et al. (2016). Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors. Journal of Biotechnology, 232, 79-88. doi:10.1016/j.jbiotec.2016.05.012
Gren, T., Ortseifen, V., Wibberg, D., Schneiker-Bekel, S., Bednarz, H., Niehaus, K., Zemke, T., Persicke, M., Pühler, A., and Kalinowski, J. (2016). Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors. Journal of Biotechnology 232, 79-88.
Gren, T., et al., 2016. Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors. Journal of Biotechnology, 232, p 79-88.
T. Gren, et al., “Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors”, Journal of Biotechnology, vol. 232, 2016, pp. 79-88.
Gren, T., Ortseifen, V., Wibberg, D., Schneiker-Bekel, S., Bednarz, H., Niehaus, K., Zemke, T., Persicke, M., Pühler, A., Kalinowski, J.: Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors. Journal of Biotechnology. 232, 79-88 (2016).
Gren, Tetiana, Ortseifen, Vera, Wibberg, Daniel, Schneiker-Bekel, Susanne, Bednarz, Hanna, Niehaus, Karsten, Zemke, Till, Persicke, Marcus, Pühler, Alfred, and Kalinowski, Jörn. “Genetic engineering in Actinoplanes sp SE50/110-development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors”. Journal of Biotechnology 232 (2016): 79-88.

2 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

The MalR type regulator AcrC is a transcriptional repressor of acarbose biosynthetic genes in Actinoplanes sp. SE50/110.
Wolf T, Droste J, Gren T, Ortseifen V, Schneiker-Bekel S, Zemke T, Pühler A, Kalinowski J., BMC Genomics 18(1), 2017
PMID: 28743243

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