A number of gene clusters carry a speci c transcrip tional activator gene which is embedded within the cluster. To find out the boundaries in the novel gene cluster and also to discriminate the impact on secondary metabolism, we constructed strains overexpressing the putative TF encoding gene dbaA or dbaG, respectively, underneath the manage within the inducible nitrate re ductase gene promoter. The overexpression Cilengitide 188968-51-6 of dbaG led to no signi cant improvements in phe notype, whereas the overexpression of dbaA brought on a strong ex tracellular pigmentation and also a reduced development diameter with the colony. Interestingly, the pigmentation depends on pH and it is reversible, in neutral and standard milieus, the culture ltrate was yellow, while at a pH of three, it turned colorless. We performed Northern hybridization experiments together with the dbaA overexpressing and dbaG OE strains.
All genes starting up from AN7893 to AN7909 had been made use of as probes, where we in contrast the promoter repressing and induc ing circumstances for the corresponding TF. The dbaG overexpress ing strain exhibited enhanced expression of the putative oxi doreductase selleck chemicals pf-562271 gene dbaF only, whereas the expression ranges in the AN7893, dbaA, dbaC, and dbaD genes even decreased. In contrast, the overexpression of dbaA coordinately upregu lated all consecutive genes in the AN7897 gene for the PKS encoding AN7903 gene,indicating that these genes kind a cluster which is managed from the fungal Zn two Cys6 TF DbaA, encoded through the most five upstream situated gene. DbaA also controls the second putative TF gene, dbaG,suggesting a complex transcriptional con trol on the complete dba gene cluster. By comparisons of your intergenic areas from the dba cluster,a motif shared by all ve sequences was identified that is not present inside the intergenic areas within the neigh boring genes except to the intergenic area of AN7893/7894.
A regulation for AN7893 and AN7894 was also de tected during the transcriptome data but not by Northern hybridization. The shared motif exhibits signi cant similarities towards the binding websites with the yeast Zn two Cys6 TFs RGT1 and

ECM22,corroborating our ndings. DHMBA and DHPDI are mutually unique metabolites. In order to identify the SMs developed by the dba gene cluster, wild kind and dbaA OE strains had been compared after cultivation in pro moter inducing medium. Culture ltrates have been extracted with ethyl acetate and subsequently analyzed by substantial efficiency liq uid chromatography coupled by using a UV diode array detector. The examination exposed a serious peak in the ten. 3 min retention time with absorption maxima at 221 and 296 nm to the dbaA OE strain and a peak at a 10. 6 min retention time with absorption maxima at 231 and 276 nm to the wild kind strain. Interestingly, the two peaks had been mutually unique.