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

doi:10.1023/A:1008891106662

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 journal › Article › peer-review

2 Citations (Scopus)

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 language	English
Pages (from-to)	15-19
Number of pages	5
Journal	Biotechnology Techniques
Volume	12
Issue number	1
DOIs	https://doi.org/10.1023/A:1008891106662
Publication status	Published - 1998

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

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.",

author = "J{\o}rgensen, \{Steen T.\} and Martin Tangney and J{\o}rgensen, \{P. L.\} and B{\o}rge Diderichsen",

year = "1998",

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language = "English",

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pages = "15--19",

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T1 - Integration and amplification of a cyclodextrin glycosyltransferase gene from Thermoanaerobacter sp. ATCC 53627 on the Bacillus subtilis chromosome

AU - Jørgensen, Steen T.

AU - Tangney, Martin

AU - Jørgensen, P. L.

AU - Diderichsen, Børge

PY - 1998

Y1 - 1998

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/0031889283

U2 - 10.1023/A:1008891106662

DO - 10.1023/A:1008891106662

M3 - Article

SN - 0951-208X

VL - 12

SP - 15

EP - 19

JO - Biotechnology Techniques

JF - Biotechnology Techniques

IS - 1

ER -

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

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