Project

Part:BBa_K1314013

Designed by: Seak Chi U, Chan Wai Lun, CHIU, Yee Ting   Group: iGEM14_Hong_Kong-CUHK   (2014-10-07)

nifK-XhoI-nifH promoter T7 expression system(strong)

homologous integration knock out nitrogenase gene with nifH promoter T7 expression system

Usage and Biology

Ammonium repleted condition(left) versus nitrogen depleted condition(right) for constitutive RFP expression and nifH regulated RFP expression in A. vinelandii

Under high Ammonium level, there is strong expression of RFP under constitutive promoter. Under nitrogen depleted condition, there is no growth of A.vinelandii. This proves that the stable genome integration is successful. The mutated A.vinelandii has already lost nitrogenase gene, and thus no longer be diazotrophic.

EcoRI PstI double digestion checking our biobrick, it shows a band of insert with expected size of 5000 bp.



Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3281
    Illegal XhoI site found at 171
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 4023
    Illegal AgeI site found at 4135
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None