Journal: Microbial Biotechnology
Article Title: The biosynthetic genes for prenylated phenazines are located at two different chromosomal loci of Streptomyces cinnamonensis DSM 1042
Figure Lengend Snippet: A. Scheme of the gene inactivation of epzP . aac(3)IV , apramycin resistance gene; Paac, promoter of the apramycin resistance gene; FRT, FLP recognition target; ori T, origin of transfer from RK2; neo , kanamycin resistance gene. Out of scale. B. Confirmation of the genotype of single‐cross‐over mutants (SCO) and double‐cross‐over mutants (DCO). Lane M: marker, C1: cosmid 8‐4D (wild type), C2: cosmid epz KG03 (Δ epzP ). Lanes 4, 6 and 7: SCO mutants. Lanes 1, 2 and 5: DCO mutants. C. HPLC/MS analysis of the formation of endophenazine A, endophenazine B and phenazine‐1‐carboxylic acid (PCA) in S. cinnamonensis DSM 1042 (wild type), the Δ epzP mutant and the Δ epzP mutant complemented with intact epzP .
Article Snippet: Chemicals and enzymes Carbenicillin and kanamycin were purchased from GenAxxon BioSciences GmbH, Biberach, Germany; apramycin, nalidixic acid and sodium persulfate (Na2 S2 O8 ) from Sigma‐Aldrich, Steinheim, Germany; chloramphenicol and sodium dithionite (Na2 S2 O4 ) from Merck, Darmstadt, Germany; and phenazine‐1‐carboxylic acid from InFarmatik, Hungary.
Techniques: Marker, High Performance Liquid Chromatography, Mass Spectrometry, Mutagenesis