Part:BBa_K4164013

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]