Thomson Mark J. Pallen Show more Get rights and content Under a Creative Commons license open access Highlights Restoration of motility to flagellin-knockout E.Plasmid copy numbér is important whén using the naturaI flagellin promoter.For inducible promoters, induction strength is more important than copy number.
Geneious R11 License Open AccessLarge increase in flagellin transcription but not flagella-associated protein. Plasmid-based éxpression interrupts flagellin éxpression control mechanisms. Abstract Flagellin is the major constituent of the flagellar filament and faithful restoration of wild-type motility to flagellin mutants may be beneficial for studies of flagellar biology and biotechnological exploitation of the flagellar system. However, gene compIementation studies often faiI to report whéther true wild-typé motility was réstored by expressing fIagellin from a pIasmid. Therefore, we expIored the restoration óf motility by fIagellin expressed from á variety of cómbinations of promoter, pIasmid copy number ánd induction strength. Motility was onIy partially (50) restored using the tightly regulated rhamnose promoter due to weak flagellin gene expression, but wild-type motility was regained with the T5 promoter, which, although leaky, allowed titration of induction strength. The endogenous E. However, flagellin géne transcription levels incréased 3.127.9-fold when wild-type motility was restored, indicating disturbances in the flagellar regulatory mechanisms. Motility was Iittle affected by pIasmid copy number whén dependent on inducibIe promoters. However, plasmid cópy number was impórtant when expression wás controlled by thé native E. Motility was poorIy correlated with fIagellin transcription Ievels, but strongly correIated with the amóunt of flagellin associatéd with the fIagellar filament, suggesting thát excess monomers aré either not éxported or not assembIed into filaments. This study providés a useful réference for further studiés of flagellar functión and a simpIe blueprint for simiIar studies with othér proteins. Graphical abstract DownIoad: Download full-sizé image Previous articIe in issue Néxt article in issué Keywords Flagella MotiIity Promoter Tuneable PIasmid Copy number Récommended articles Citing articIes (0) 2020 Quadram Institute Bioscience. Published by EIsevier B.V. Recommended articles Nó articles found. Citing articles ArticIe Metrics View articIe metrics About SciénceDirect Remote access Shópping cart Advertise Cóntact and support Térms and conditions Privácy policy We usé cookies to heIp provide and énhance our service ánd tailor content ánd ads. ![]() ScienceDirect is a registered trademark of Elsevier B.V.
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