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

Steen T. Jørgensen, Martin Tangney, P. L. Jørgensen, Børge Diderichsen

    Research output: Contribution to journalArticle

    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.

    Original languageEnglish
    Pages (from-to)15-19
    Number of pages5
    JournalBiotechnology Techniques
    Volume12
    Issue number1
    DOIs
    Publication statusPublished - 1998

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    Thermoanaerobacter
    Bacillus subtilis
    chromosomes
    genes
    plasmids
    cyclomaltodextrin glucanotransferase
    DNA

    Cite this

    Jørgensen, Steen T. ; Tangney, Martin ; Jørgensen, P. L. ; Diderichsen, Børge. / Integration and amplification of a cyclodextrin glycosyltransferase gene from Thermoanaerobacter sp. ATCC 53627 on the Bacillus subtilis chromosome. In: Biotechnology Techniques. 1998 ; Vol. 12, No. 1. pp. 15-19.
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    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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    Integration and amplification of a cyclodextrin glycosyltransferase gene from Thermoanaerobacter sp. ATCC 53627 on the Bacillus subtilis chromosome. / Jørgensen, Steen T.; Tangney, Martin; Jørgensen, P. L.; Diderichsen, Børge.

    In: Biotechnology Techniques, Vol. 12, No. 1, 1998, p. 15-19.

    Research output: Contribution to journalArticle

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    AU - Diderichsen, Børge

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