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Integration and amplification of a cyclodextrin glycosyltransferase gene from Thermoanaerobacter sp. ATCC 53627 on the Bacillus subtilis chromosome

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Biotechnology Techniques

Abstract

A plasmid was constructed containing the replication functions of pUC19, and the cgtA gene from Thermoanaerobacter sp. ATCC 53627, flanked by the dal gene from Bacillus subtilis and a sequence downstream from this gene. This was transformed into a Dal- B. subtilis strain, selecting for Dal+ transformants, which contained the cgtA gene in single copy integrated in the B. subtilis chromosome. The gene was subsequently amplified by a method which ensured that there was no functional plasmid replication system on the integrated DNA. The amplified structure was stable in the absence of selection pressure.

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Authors
  1. Steen T. Jorgensen
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  2. Martin Tangney
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  3. Per Lina Jorgensen
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  4. Borge Diderichsen
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Jorgensen, S.T., Tangney, M., Jorgensen, P.L. et al. Integration and amplification of a cyclodextrin glycosyltransferase gene from Thermoanaerobacter sp. ATCC 53627 on the Bacillus subtilis chromosome. Biotechnology Techniques 12, 15–19 (1998). https://doi.org/10.1023/A:1008891106662

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  • DOI: https://doi.org/10.1023/A:1008891106662

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