A versatile system consisting of an integrational vector and a bacitracin (Bt)-producing β-galactosidase (β-Gal)-negative (Lac) TLH strain was constructed to quantify promoter activity and to study gene regulation in a single-copy set-up. The vector pTLH utilizes the promoterless gene derived from pQF52 and contains the pBR322 origin of replication and a kanamycin-resistance gene for selection in both and The vector also contains an inner part of the first gene of the Bt synthetase operon which enables its integration into the bts of by Campbell-type recombination. This recombination event can be easily tested on a lawn where loss of Bt production, i.e. no clearing zone on the lawn, is indicative of the proper integration. The Lac TLH strain was developed by elimination of the natural β-Gal activity of strain ATCC 10716 UM12 using NTG mutagenesis.


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