title: TREX: a universal tool for the transfer and expression of biosynthetic pathways in bacteria.
abstract: Secondary metabolites represent a virtually inexhaustible source of natural molecules exhibiting a high potential as pharmaceuticals or chemical building blocks. To gain broad access to these compounds, sophisticated expression systems are needed that facilitate the transfer and expression of large chromosomal regions, whose genes encode complex metabolic pathways. Here, we report on the development of the novel system for the transfer and expression of biosynthetic pathways (TREX), which comprises all functional elements necessary for the delivery and concerted expression of clustered pathway genes in different bacteria. TREX employs (i) conjugation for DNA transfer, (ii) randomized transposition for its chromosomal insertion, and (iii) T7 RNA polymerase for unimpeded bidirectional gene expression. The applicability of the TREX system was demonstrated by establishing the biosynthetic pathways of two pigmented secondary metabolites, zeaxanthin and prodigiosin, in bacteria with different metabolic capacities. Thus, TREX represents a valuable tool for accessing natural products by allowing comparative expression studies with clustered genes.
authors: Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf, Research Center Jülich, Jülich, Germany.
authors: Anita
authors: A
authors: Loeschcke
authors: Annette
authors: A
authors: Markert
authors: Susanne
authors: S
authors: Wilhelm
authors: Astrid
authors: A
authors: Wirtz
authors: Frank
authors: F
authors: Rosenau
authors: Karl-Erich
authors: KE
authors: Jaeger
authors: Thomas
authors: T
authors: Drepper
mesh_terms: D001419:Bacteria / Q000235:genetics* / Q000378:metabolism*; D053898:Biosynthetic Pathways; D002876:Chromosomes, Bacterial; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D011353:Prodigiosin / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D024341:Xanthophylls / Q000235:genetics; D065146:Zeaxanthins
chemical_list: D014764:Viral Proteins; D024341:Xanthophylls; D065146:Zeaxanthins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D011353:Prodigiosin