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@ARTICLE{Weihmann:866203,
      author       = {Weihmann, Robin and Domröse, Andreas and Drepper, Thomas
                      and Jaeger, Karl‐Erich and Loeschcke, Anita},
      title        = {{P}rotocols for y{TREX} /{T}n5‐based gene cluster
                      expression in {P}seudomonas putida},
      journal      = {Microbial biotechnology},
      volume       = {13},
      number       = {1},
      issn         = {1751-7915},
      address      = {Oxford},
      publisher    = {Wiley-Blackwell},
      reportid     = {FZJ-2019-05374},
      pages        = {250-262},
      year         = {2020},
      abstract     = {Bacterial gene clusters, which represent a genetic treasure
                      trove for secondary metabolite pathways, often need to be
                      activated in a heterologous host to access the valuable
                      biosynthetic products. We provide here a detailed protocol
                      for the application of the yTREX ‘gene cluster
                      transplantation tool’: Via yeast recombinational cloning,
                      a gene cluster of interest can be cloned in the yTREX
                      vector, which enables the robust conjugational transfer of
                      the gene cluster to bacteria like Pseudomonas putida, and
                      their subsequent transposon Tn5‐based insertion into the
                      host chromosome. Depending on the gene cluster architecture
                      and chromosomal insertion site, the respective pathway genes
                      can be transcribed effectively from a chromosomal promoter,
                      thereby enabling the biosynthesis of a natural product. We
                      describe workflows for the design of a gene cluster
                      expression cassette, cloning of the cassette in the yTREX
                      vector by yeast recombineering, and subsequent transfer and
                      expression in P. putida. As an example for yTREX‐based
                      transplantation of a natural product biosynthesis, we
                      provide details on the cloning and activation of the
                      phenazine‐1‐carboxylic acid biosynthetic genes from
                      Pseudomonas aeruginosa in P. putidaKT2440 as well as the use
                      of β‐galactosidase‐encoding lacZ as a reporter of
                      production levels.},
      cin          = {IMET / IBG-1},
      ddc          = {610},
      cid          = {I:(DE-Juel1)IMET-20090612 / I:(DE-Juel1)IBG-1-20101118},
      pnm          = {581 - Biotechnology (POF3-581)},
      pid          = {G:(DE-HGF)POF3-581},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:31162833},
      UT           = {WOS:000596580200022},
      doi          = {10.1111/1751-7915.13402},
      url          = {https://juser.fz-juelich.de/record/866203},
}