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Post-transcriptional Regulation of RsmA In Pseudomonas aeruginosaMiller, Ian 01 August 2018 (has links) (PDF)
Pseudomonas aeruginosa is a Gram-negative bacillus found in numerous environments. Gene regulatory mechanisms such as; Two-Component Systems, transcriptional and post-transcriptional regulators, and small non-coding RNAs control the expression of virulence factors that allow P. aeruginosa to initiate acute infections and persist as a chronic infection. A significant post-transcriptional regulator involved in these regulatory networks is the Regulator of Secondary Metabolites (RsmA). In this study, we investigated the contribution of a putative stem-loop on expression of RsmA. We constructed rsmA leader fusions to measure translation with and without the stem-loop present. Secondly, we introduced point mutations to disrupt the formation of the stem-loop. Finally, we performed Site-Directed Mutagenesis on the rsmA leader to examine protein levels in vivo by western blot analysis using an HA-tagged rsmA. Our data suggests that the segment of RNA that contains the putative stem-loop structure serves some function in post-transcriptional regulation of RsmA.
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