Generator

Part:BBa_K4202007

Designed by: LIAOLIAO GAO   Group: iGEM22_ZJU-China   (2022-09-25)
Revision as of 03:42, 26 September 2022 by SinubilovesCola (Talk | contribs)

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PHY-Oxylimited-SpaBCTS

The subtilisin production module is mediated by a hypoxic promoter and functions through a gene cluster composed of SpaBCTS. The population of engineered Bacillus subtilis keeps growing in the cracks, consuming oxygen, and at the same time, the engineered bacteria penetrate deep into the tiny cracks, and the environment lacks oxygen, thus realizing hypoxic conditions, and the subtilisin production module is activated. However we failed to convince the function as the bacteria could only express the first protein under oxygen limitation environment.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal BamHI site found at 2495
    Illegal BamHI site found at 4765
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
    Illegal AgeI site found at 2309
    Illegal AgeI site found at 3317
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 361
    Illegal SapI.rc site found at 2409
    Illegal SapI.rc site found at 4919
    Illegal SapI.rc site found at 6121


Applications of BBa_K4202007

we transformed it into our chassis-Bacillus subtilis for further protein expression. 50ml shaking flasks with 5ml culture media were introduced to reach an extra-large bacterial population while the normal 12ml shaking bacteria tubes were used as control. From the SDS-PAGE image, we could see a clear strip locating in 120kDa’s area for the 50ml shaking flasks group, revealing the excellent function of Oxygen-limitation induced promotor. However, it seemed that only the first gene (SpaB,120kDa) had expressed under the condition, suggesting that the original sequence didn’t match well to the promotor.

GLL-5.png

Fig 1-1Characterization of Oxygen-limitation induced promotor by SDS-PAGE

See the [1] ZJU-China 2022 wiki for more details.

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