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| 1 | Rapid generation of recombinant Pseudomonas putida secondary metabolite producers using yTREX显示文摘Microbial secondary metabolites represent a rich source of valuable compounds with a variety of applications in medicine or agriculture.Effective exploitation of this wealth of chemicals requires the functional expression of the respective biosynthetic genes in amenable heterologous hosts.We have previously established the TREX system which facilitates the transfer,integration and expression of biosynthetic gene clusters in various bacterial hosts.Here,we describe the yTREX system,a new tool adapted for one-step yeast recombinational cloning of gene clusters.We show that with yTREX,Pseudomonas putida secondary metabolite production strains can rapidly be constructed by random targeting of chromosomal promoters by Tn5 transposition.Feasibility of this approach was corroborated by prodigiosin production after yTREX cloning,transfer and expression of the respective biosynthesis genes from Serratia marcescens.Furthermore,the applicability of the system for effective pathway rerouting by gene cluster adaptation was demonstrated using the violacein biosynthesis gene cluster from Chromobacterium violaceum,producing pathway metabolites violacein,deoxyviolacein,prodeoxyviolacein,and deoxychromoviridans.Clones producing both prodigiosin and violaceins could be readily identified among clones obtained after random chromosomal integration by their strong color-phenotype.Finally,the addition of a promoter-less reporter gene enabled facile detection also of phenazine-producing clones after transfer of the respective phenazine-1-carboxylic acid biosynthesis genes from Pseudomonas aeruginosa.All compounds accumulated to substantial titers in the mg range.We thus corroborate here the suitability of P.putida for the biosynthesis of diverse natural products,and demonstrate that the yTREX system effectively enables the rapid generation of secondary metabolite producing bacteria by activation of heterologous gene clusters,applicable for natural compound discovery and combinatorial biosynthesis. | Andreas Domrose Robin Weihmann Stephan Thies Karl-Erich Jaeger Thomas Drepper Anita Loeschcke | 2017 | Synthetic and Systems Biotechnology2017，2，4： | 2 |
| 2 | Pathogenic Cycle Between the Endogenous Nitric Oxide Synthase Inhibitor Asymmetrical Dimethylarginine and the Leukocyte-Derived Hemoprotein Myeloperoxidase显示文摘 | Eike-Christin von Leitner Anna Klinke Dorothee Atzler Jessica L. Slocum Natalie Lund Jan T. Kielstein Renke Maas Robin Schmidt-Haupt Michaela Pekarova Olaf Hellwinkel Dimitrios Tsikas Louis G. D?Alecy Denise Lau Stephan Willems Lukas Kubala Heimo Ehmke Th | 2011 | Circulation2011，，24： | 1 |
| 3 | National science training policy and early scientific careers in france and the united states 显示文摘 | Monica Ganghan Stephane Robin | 2004 | Research Policy2004，，33： | 1 |
| 4 | Pathogenesis of Varicose Veins显示文摘 | Rahmi Oklu Roy Habito Manuel Mayr Amy R. Deipolyi Hassan Albadawi Robin Hesketh T. Gregory Walker Katy R. Linskey Chandler A. Long Stephan Wicky Julianne Stoughton Michael T. Watkins | 2012 | Journal of Vascular and Interventional Radiology2012，，1： | 1 |
| 5 | Cooperation with pub- lic research institutions and success in innovation: Evidence from France and Germany 显示文摘 | STEPHANE ROBIN TORBEN SCHUBERT | 2013 | Research Policy2013，42，1： | 1 |
| 6 | National Science Training Policy and Early Scientific Careers in France and the United States 显示文摘 | Monica Gaughan Stephane Robin | 2004 | Research Policy2004，33，4： | 1 |
| 7 | Doconsumers really refuse to buy genetically modified food?显示文摘 | CHARLES NOUSSAIR STEPHANE ROBIN BERNARD RUFFIEUX | 2004 | The Eco-nomic Journal2004，114，492： | 1 |
| 8 | Mapping vulnerability to multiple stressors: climate change and globalization in India显示文摘 | Karen O’Brien Robin Leichenko Ulka Kelkar Henry Venema Guro Aandahl Heather Tompkins Akram Javed Suruchi Bhadwal Stephan Barg Lynn Nygaard Jennifer West | 2004 | Global Environmental Change2004，，： | 1 |
| 9 | A Set of SAS Macros for the Analysis of Variance of Gene Expression Data 显示文摘 | Christelle Hennequet-Antier Helene Chiapello Karine Piot Severine Degrelle Isabelle Hue Jean-Paul Renard Francois Rodolphe Stephane Robin | 2005 | BMC Bioinformatics2005，，6： | 1 |
| 10 | Do consumers not about biotech foods or do they just not read the labels?显示文摘 | Charles Noussair Stephane Robin Bernard Ruffieux | 2002 | Economics Letters2002，，75： | 1 |
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