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

Paper ID Volume ID Publish Year Pages File Format Full-Text
22538 43366 2016 10 PDF Available
Title
Genetic engineering in Actinoplanes sp. SE50/110 − development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors
Abstract

•Basic genetic engineering toolkit established for Actinoplanes sp. SE50/110.•Development of the Actinoplanes sp. SE50/110 - E. coli intergeneric conjugation.•Integration sites for ϕC31, ϕBT1 and VWB actinophage-based integration systems sequenced and described for the first time in the Actinoplanes genus.•GUS reporter system applied successfully for Actinoplanes sp. SE50/110.

The α-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 ϕC31, ϕBT1 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 ϕC31 based vector pSET152, the integration site is highly conserved, while for ϕBT1 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.

Keywords
Acarbose; Actinoplanes sp. SE50/110; Genetic engineering; Integrative vectors
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Genetic engineering in Actinoplanes sp. SE50/110 − development of an intergeneric conjugation system for the introduction of actinophage-based integrative vectors
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 232, 20 August 2016, Pages 79–88
Authors
, , , , , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us