Part:BBa_K1963010
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. 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. 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 101
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