Part:BBa_K3137014
responce circuit
Usage and Biology
The responce gene circuit is composed of an inducible promoter PputA(BBa_K3137002) and anti-phage protein genes of the incubation period (about 5 min), and an inducible promoter PglcF (BBa_K3137003) and a toxic protein gene protegrin-1(BBa_K62800) of the burst period (about 20 min).
When E. coli is infected by a phage, responce gene circuit is induced to express. When a phage infects E. coli into the incubation period (about 5 min), the inducible promoter PputA will induce the bacteria to express the resistant proteins (BBa_K3137006)(BBa_K3137005).
If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage infection, the inducible promoter PglcF will activate the expression of downstream toxic protein gene so that the bacteria will lyse cells before the assembly of phages.
We performed the phage infection assay on the LB agar plate, and got very good resistance.
In addition, we inoculated E. coli BL21 and recombinant E. coli BL21-pET28a-PputA-abpAB-gntR-PglcF-P-1 in LB liquid medium to raise the logarithmic growth phase, i.e. OD 0.6-0.8 . Then the fresh phage solution was inoculated at the same time and culture was continued for 1-2 h. As a result, it was found that the recombinant grew well.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1280
Illegal EcoRI site found at 2048
Illegal PstI site found at 1313
Illegal PstI site found at 2705
Illegal PstI site found at 3785 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1280
Illegal EcoRI site found at 2048
Illegal PstI site found at 1313
Illegal PstI site found at 2705
Illegal PstI site found at 3785
Illegal NotI site found at 4987 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1280
Illegal EcoRI site found at 2048
Illegal BglII site found at 1619
Illegal BglII site found at 2679
Illegal BamHI site found at 481 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1280
Illegal EcoRI site found at 2048
Illegal PstI site found at 1313
Illegal PstI site found at 2705
Illegal PstI site found at 3785 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1280
Illegal EcoRI site found at 2048
Illegal PstI site found at 1313
Illegal PstI site found at 2705
Illegal PstI site found at 3785
Illegal NgoMIV site found at 4391 - 1000COMPATIBLE WITH RFC[1000]
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