A quick and reliable method for monitoring gene expression in actinomycetes.
Amycolatopsis balhimycina DSM5908 is the producer of the vancomycin-type glycopeptide antibiotic balhimycin. Balhimycin production is controlled by transcriptional regulators and depends on different environmental factors. To analyse the regulatory network P(tba), a representative promoter of the balhimycin gene cluster, was fused to the reporter gene egfp and integrated into the genome of A. balhimycina. Balhimycin production, fluorescence intensity and the amount of tba-transcript were determined under different cultivation conditions in microtiter plates. Thus we could demonstrate that these values are directly correlated. This approach facilitates the analysis of a large number of samples in parallel and it provides the opportunity to easily define conditions to increase balhimycin production.
Chevillotte
Meike
M
Lehrstuhl für Mikrobiologie/Biotechnologie, Mikrobiologisches Institut, Universität Tübingen, Germany.
Menges
Rainer
R
Muth
Günther
G
Wohlleben
Wolfgang
W
Stegmann
Evi
E
D039903:Actinobacteria / Q000235:genetics*; D001709:Biotechnology / Q000379:methods*; D004269:DNA, Bacterial / Q000378:metabolism; D005453:Fluorescence; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D009876:Operon / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014640:Vancomycin / Q000031:analogs & derivatives / Q000096:biosynthesis
D004269:DNA, Bacterial; C087127:balhimycin; D014640:Vancomycin