Composite

Part:BBa_K1463604

Designed by: iGEM14_Glasgow   Group: iGEM14_Glasgow   (2014-10-04)
Revision as of 21:33, 17 October 2014 by GemmaMcl (Talk | contribs)

FliC with RBS and J23100 promoter

FliC with B0032 and B0034 RBS under the control of J23100 promoter.


This composite part was made by inserting synthesised oligonucleotides containing J23100 promoter (strong) and B0032 into K1463601 (fliC and B0034 RBS).


As shown in Figure 1D, this construct failed to restore swimming in knockout fliC strains. On sequencing we found this plasmid to have a mutation in the promoter, explaining this result. We failed to clone a functional J23100 promoter in front of the fliC biobrick, suggesting that strong over expression of fliC may be toxic.


Flic Motility Swarm Assay

310px-GU_Figure_1_swarm_M.png

Figure 1: FliC Swarm Motility Assays.
(A) DS941, (B) DS941 ΔfliC,
(C) DS941 ΔfliC + pSB1C3 fliC (no promoter), (D) DS941 ΔfliC + J23100 (mutant promoter) fliC,
(E) DS941 ΔfliC + J23116-fliC(1), (F) DS941 ΔfliC + J23116-fliC(2),
(G) DS941 ΔfliC + J23106-fliC(1), (H) DS941 ΔfliC + J23106-fliC(2)


GU_Figure_2_Motility_histogram.png
Figure 2 - FliC Motility Histogram



For more information on the biobrick and methods used go to http://2014.igem.org/wiki/index.php?title=Team:Glasgow/Project/Mobility_Proteins#fliC Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1306
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 383
    Illegal AgeI site found at 791
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None