Microbiology 1995, 141:1691–1705.PubMedCrossRef 66. Figurski DH, Helinski DR: Replication of an origin-containing derivative buy PR-171 of plasmid RK2 dependent on a plasmid function provided in trans . Proc Natl Acad Sci USA 1979,76(4):1648–1652.PubMedCrossRef 67. Ojangu EL, Tover A, Teras R, Kivisaar M: Effects of combination of different -10 hexamers and downstream sequences on stationary-phase-specific sigma factor sigma(S)-dependent transcription in Pseudomonas

putida . J Bacteriol 2000,182(23):6707–6713.PubMedCrossRef 68. Koch B, Jensen LE, Nybroe O: A panel of Tn 7 -based vectors for insertion of the gfp marker gene or for delivery of cloned DNA into Gram-negative bacteria at a neutral chromosomal site. J Microbiol Methods 2001,45(3):187–195.PubMedCrossRef Authors’ contributions MP and RH prepared design of experimental work. MP carried out transposon mutagenesis screen and participated in OMP analysis. AA purified OMPs and did OMP find protocol pattern analysis. HI constructed mutant strains and contributed enzyme assays. RH performed lysis assays, coordinated experimental work and wrote the manuscript. All authors participated in manuscript editing and approved the final manuscript.”
“Background Sirodesmin PL is the major phytotoxin produced by Selleck OSI-906 the

plant pathogen Leptosphaeria maculans (Desm.), the causal agent of blackleg disease of Brassica napus (canola). Sirodesmin PL has antibacterial

and antiviral properties [1] and is essential for full virulence of L. maculans on stems of B. napus [2]. This toxin is an epipolythiodioxopiperazine (ETP), a class of secondary metabolites characterised by the presence of a highly reactive disulphide-bridged dioxopiperazine ring synthesised from two amino acids (for review see [3]). The first committed step in the sirodesmin biosynthetic pathway is prenylation of tyrosine [4, 5]. As for other fungal secondary metabolites, the genes for the biosynthesis of sirodesmin PL are clustered. The sirodesmin cluster contains 18 genes that are co-ordinately regulated with timing consistent with sirodesmin PL production. Disruption of one of see more these genes, sirP, which encodes a peptide synthetase, results in an isolate unable to produce sirodesmin PL [6]. Based on comparative genomics, the cluster of genes in Aspergillus fumigatus responsible for the biosynthesis of another ETP, gliotoxin, was then predicted. The pattern of expression of the clustered homologs was consistent with gliotoxin production [7]. The identity of this gene cluster was confirmed via the disruption of peptide synthetase, gliP whereby the resultant mutant was unable to make gliotoxin [8, 9]. These ETP gene clusters also encode a Zn(II)2Cys6 transcription factor, namely SirZ for sirodesmin, and GliZ for gliotoxin [7].