title: Effect of Genome Position on Heterologous Gene Expression in Bacillus subtilis: An Unbiased Analysis.
abstract: A fixed gene copy number is important for the in silico construction of engineered synthetic networks. However, the copy number of integrated genes depends on their genomic location. This gene dosage effect is rarely addressed in synthetic biology. Two studies in Escherichia coli presented conflicting data on the impact of gene dosage. Here, we investigate how genome location and gene orientation influences expression in Bacillus subtilis. An important difference with the E. coli studies is that we used an unbiased genome integration approach mediated by random transposon insertion. We found that there is a strong gene dosage effect in fast growing B. subtilis cells, which can amount to a 5-fold difference in gene expression. In contrast, gene orientation with respect to DNA replication direction does not influence gene expression. Our study shows that gene dosage should be taken into account when designing synthetic circuits in B. subtilis and presumably other bacteria.
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University , Richardson Road, NE2 4AX Newcastle, United Kingdom.|DSM Biotechnology Center , P.O. Box 1, 2600 MA Delft, The Netherlands.
authors.forename: Christopher
authors.initials: C
authors.lastname: Sauer
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University , Richardson Road, NE2 4AX Newcastle, United Kingdom.
authors.forename: Simon
authors.initials: S
authors.lastname: Syvertsson
authors.affiliation: Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam , De Boelelaan, 1081 HZ Amsterdam, The Netherlands.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Bohorquez
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University , Richardson Road, NE2 4AX Newcastle, United Kingdom.|DSM Biotechnology Center , P.O. Box 1, 2600 MA Delft, The Netherlands.
authors.forename: Rita
authors.initials: R
authors.lastname: Cruz
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University , Richardson Road, NE2 4AX Newcastle, United Kingdom.
authors.forename: Colin R
authors.initials: CR
authors.lastname: Harwood
authors.affiliation: DSM Biotechnology Center , P.O. Box 1, 2600 MA Delft, The Netherlands.
authors.forename: Tjeerd
authors.initials: T
authors.lastname: van Rij
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University , Richardson Road, NE2 4AX Newcastle, United Kingdom.|Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam , De Boelelaan, 1081 HZ Amsterdam, The Netherlands.
authors.forename: Leendert W
authors.initials: LW
authors.lastname: Hamoen
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D004261:DNA Replication / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics*; D058615:Synthetic Biology / Q000379:methods
keywords: Bacillus subtilis; GFP; LacZ; gene expression; genome location; transposon