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. 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
- 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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