Difference between revisions of "Part:BBa K1963010"

Revision as of 12:54, 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>/i> hairpin, followed by the strong terminator sequence T1/TE. The anti RBS-<i>fliC sRNA is placed after C-193. The system should impair motility of E. coli as a proof-of-concept.

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.

Experimental procedures needed.

Figure 1. Bar graph showing average colony area and standard deviation of replicates. Photographs representative of motility assays with BBa_K1963002, BBa_K1963004 and BBa_K1963010.

Sequence and Features