2021
DOI: 10.21203/rs.3.rs-944639/v1
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The Sequence of the Ribosomal Binding Site Controls the Gene Expression in Brucella

Abstract: BackgroundBrucella is an important pathogen causing Brucellosis. Vaccine strains obtained by a single knockout cannot combine low virulence and immunogenicity. Our study modified the SD sequence and spacer sequence of the RBS of Brucella to affect its protein expression. We altered the RBS of LPS-associated genes to reduce LPS-associated protein expression while retaining LPS integrity.ResultsWe first established an evaluation system based on the reporter gene red fluorescent protein mCherry. The mCherry expre… Show more

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Cited by 2 publications
(3 citation statements)
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“…Or the spacer between Shine-Dalgarno (SD) sequence and start codon make the different results. The nucleotide composition among the SD sequence and start codon has important impact on the expression [43][44][45]. But there has few research about these two promoters, it is hard to de ne the exact point that cause the difference.…”
Section: Discussionmentioning
confidence: 99%
“…Or the spacer between Shine-Dalgarno (SD) sequence and start codon make the different results. The nucleotide composition among the SD sequence and start codon has important impact on the expression [43][44][45]. But there has few research about these two promoters, it is hard to de ne the exact point that cause the difference.…”
Section: Discussionmentioning
confidence: 99%
“…Various technologies have been developed as powerful tool sets for manipulating gene expression. While fine-tuning transcriptional regulators (i.e., promoters) has cemented their importance in the synthetic toolbox, other metabolic engineering tactics popular for its ability to directly tune protein synthesis levels within a pathway are ribosomal binding site (RBS) variation and RBS accessibility. , For example, directly tuning each enzyme in the pathway of interest by cis -mediated riboregulation (that act by occluding the RBS) has proven to reduce cellular burden and optimize titer, yield, and biomass in various metabolic engineering strategies (reviewed by Kent et al). In this way, a bioengineer can manipulate protein production within a metabolic engineering pathway by directly tuning down specific enzymes at the protein level, rather than at a mRNA transcript level, thus allowing for an almost digitally precise fine-tuning approach. ,,,,, Additionally, because of the cis -acting function, there is no risk of off-target effects and no need to balance regulatory stoichiometry.…”
Section: Introductionmentioning
confidence: 99%
“…However, existing RBS occlusion tools may suffer from an unpredictable dynamic range and lack of design rules for achieving optimal performance . Finally, the same RBS sequence in different genetic backgrounds has been shown to lead to large differences in protein production levels. , Thus, there is a need to develop a more ‘portable’ design across not only genetic contexts but also coding sequence contexts. Here, we describe the development of tight protein production control that is evident in two different promoter/reporter sequence contexts, as well as different genetic backgrounds.…”
Section: Introductionmentioning
confidence: 99%