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Part:BBa_K314982:Design

Designed by:   Group: iGEM10_Washington   (2010-10-07)
Revision as of 05:17, 7 October 2010 by Eljefe (Talk | contribs) (→‎References)

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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
    COMPATIBLE WITH RFC[1000]


Design Notes

All the essential genes for our T6SS are contained within two putative operons, encoded in opposite directions. The native promoters for both operons are found in the same intergenic region, between fha1 and tssA1. Therefore, we can easily replace the promoter regions for both operons in one step.


Source

In order to create a probiotic application for this system, we first attempt to express it heterologously in non-pathogenic E. Coli. Starting from a Fosmid containing our T6SS, we are using Recombineering to replace the strict native regulation with robust T7 promoters to create strong expression of the T6SS.

References

mention that fosmid contains natural promoter, may not be trascribed from at all, or at right amount for a plasmid ( more than 1 copy of plasmid)