Composite

Part:BBa_K4164013

Designed by: Bo Cheng   Group: iGEM22_NAU-CHINA   (2022-09-30)


Inductive expression of BSS

In order to find an appropriate expression intensity to achieve balance between metabolic burden and detection efficiency, we also tried T7lac promoter from (pET-29a+)(Novagen).

We used the methods described by Y. TAZUKE et al. (1992) to assay sulfatase activity. Figure 5 shows that the activity of BSS was estimated at approximately 0.15 unit.


Fig.1 Assay of sulfatase activity. A reaction mixture containing 1.0 ml of 100 mM Tris buffer (pH 8.0), 0.20 ml of 15mM [β-NAD, 0.20ml of 1mM CDCA-S, and 1.45ml of distilled water in eppendorf tube was incubated at 30°C for 10min, and then added 0.1ml of the cell-free solution and 0.05 ml of β-HSD solution (10 units/ml).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1600
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1001
    Illegal NgoMIV site found at 1052
  • 1000
    COMPATIBLE WITH RFC[1000]


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