PT  - JOURNAL ARTICLE
AU  - Joep Houkes
AU  - Lorenzo Olivi
AU  - Zacharie Paquet
AU  - Nico J. Claassens
AU  - John van der Oost
TI  - Design, construction and optimization of a synthetic RNA polymerase operon in <em>Escherichia coli</em>
AID  - 10.1101/2021.11.05.467461
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.11.05.467461
4099  - http://biorxiv.org/content/early/2021/11/05/2021.11.05.467461.short
4100  - http://biorxiv.org/content/early/2021/11/05/2021.11.05.467461.full
AB  - Prokaryotic genes encoding functionally related proteins are often clustered in operons. The compact structure of operons allows for co-transcription of the genes, and for co-translation of the polycistronic messenger RNA to the corresponding proteins. This leads to reduced regulatory complexity and enhanced gene expression efficiency, and as such to an overall metabolic benefit for the protein production process in bacteria and archaea. Interestingly, the genes encoding the subunits of one of the most conserved and ubiquitous protein complexes, the RNA polymerase, are not clustered in a single operon. Rather, its genes are scattered in all known prokaryotic genomes, generally integrated in different ribosomal operons. To analyze the impact of this genetic organization on the fitness of Escherichia coli, we constructed a bacterial artificial chromosome harboring the genes encoding the RNA polymerase complex in a single operon. Subsequent deletion of the native chromosomal genes led to a reduced growth on minimal medium. However, by using adaptive laboratory evolution the growth rate was restored to wild-type level. Hence, we show that a highly conserved genetic organization of core genes in a bacterium can be reorganized by a combination of design, construction and optimization, yielding a well-functioning synthetic genetic architecture.Competing Interest StatementThe authors have declared no competing interest.