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
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BamHI site found at 3281
Illegal XhoI site found at 171
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 4023
Illegal AgeI site found at 4135
- 1000COMPATIBLE WITH RFC