Difference between revisions of "Part:BBa K1963010"

Latest revision as of 15:20, 23 October 2016

An sRNA system for down-regulation of E. coli fliC encoding flagellin

This is a multi-sequence system for small RNA (sRNA) production in an E. coli host - or especially with co-expression of E. coli Hfq BBa_K1963000. The system is based on the BBa_K1963002 template and has the proD strong promoter, followed by a short antisense sequence targeting the RBS and 5' end of the E. coli fliC transcript, followed by the E. coli micC hairpin, followed by the strong terminator sequence T1/TE. The anti RBS-fliC sRNA is placed after C-193. The system should impair motility of E. coli as a proof-of-concept.

Note that this part differs from BBa_K1963004 slightly in that the sRNA for this one covers the fliC RBS

Usage and Biology

The motile E. coli host strain MG1655 was tranformed with BBa_K1963010 and subjected to a plate-based swimming assay (Figure 1). The BBa_K1963010 contains a sRNA that will target the RBS and initial codons of the fliC transcript that encodes flagellin - the principle component of the flagellum. The data related to this biobrick are labelled spiRNA-fliC-RBS-CDS in Figure 1.

Experimental procedures needed.

Figure 1. sRNA Devices can Inhibit Swimming in an E. coli Test System. Photographs representative of motility assays with MG1655 transformed with BBa_K1963002 (control), BBa_K1963004 (spiRNA-fliC-CDS)and BBa_K1963010 (spiRNA-fliC-RBS-CDS). Bar graph showing average swimming/spreading area and standard deviation of replicates.

Sequence and Features