RNA

Part:BBa_K1963004

Designed by: Frank Sargent   Group: iGEM16_Dundee_Schools   (2016-10-07)
Revision as of 13:27, 23 October 2016 by Franksargent (Talk | contribs) (Usage and Biology)

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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 5' end of the fliC transcript, followed by the E. coli micC hairpin, followed by the strong terminator sequence T1/TE. The anti fliC sRNA is placed betweeen C-193 and T-194. The system should impair motility of E. coli.

Usage and Biology

The motile E. coli host strain MG1655 was tranformed with BBa_K1963004 and subjected to a plate-based swimming assay (Figure 1). The BBa_K1963004 contains a sRNA that will target the 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-CDS in Figure 1.


experimental procedure needed.


Ecoliflicfrulhuq2.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 101


[edit]
Categories
//awards/part_collection/2016
Parameters
None