title: Combinatorial engineering for heterologous gene expression.
abstract: Tools for strain engineering with predictable outcome are of crucial importance for the nascent field of synthetic biology. The success of combining different DNA biological parts is often restricted by poorly understood factors deriving from the complexity of the systems. We have previously identified variants for different regulatory elements of the expression cassette XylS/Pm. When such elements are combined they act in a manner consistent with their individual behavior, as long as they affect different functions, such as transcription and translation. Interestingly, sequence context does not seem to influence the final outcome significantly. Expression of reporter gene bla could be increased up to 75 times at the protein level by combining three variants in one cassette. For other tested reporter genes similar results were obtained, except that the stimulatory effect was quantitatively less. Combination of individually characterized DNA parts thus stands as suitable method to achieve a desired phenotype.
authors: Department of Biotechnology; Norwegian University of Science and Technology; Trondheim, Norway.
authors: Friederike
authors: F
authors: Zwick
authors: Department of Biotechnology; Norwegian University of Science and Technology; Trondheim, Norway.
authors: Rahmi
authors: R
authors: Lale
authors: Department of Biotechnology; Norwegian University of Science and Technology; Trondheim, Norway.
authors: Svein
authors: S
authors: Valla
mesh_terms: D057005:Bioengineering / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D017930:Genes, Reporter; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D058615:Synthetic Biology; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D004247:DNA; D001618:beta-Lactamases
keywords: Escherichia coli; XylS/Pm; combinatorial engineering; heterologous gene expression; metabolic engineering; recombinant protein production; synthetic biology
references: J Biol Eng. 2008 Apr 14;2:5
references: 18410688
references: Appl Environ Microbiol. 2009 Apr;75(7):2002-11
references: 19201973
references: J Mol Biol. 1990 Jan 20;211(2):427-45
references: 2106583
references: Microb Biotechnol. 2010 Jan;3(1):38-47
references: 21255304
references: J Biol Eng. 2010 Jan 20;4(1):1
references: 20205762
references: Microb Cell Fact. 2013 Mar 18;12:26
references: 23506076
references: Plasmid. 1997;38(1):35-51
references: 9281494
references: Appl Environ Microbiol. 2011 Apr;77(8):2648-55
references: 21335387
references: Microb Biotechnol. 2009 May;2(3):379-89
references: 21261932
references: Appl Environ Microbiol. 2004 Dec;70(12):7033-9
references: 15574897