Difference between revisions of "Part:BBa K4164013"
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| − | In order to find an appropriate expression intensity to achieve balance between metabolic burden and detection efficiency, we also tried T7<em>lac</em> promoter from pET-29a+(Novagen). | + | In order to find an appropriate expression intensity to achieve balance between metabolic burden and detection efficiency, we also tried T7<em>lac</em> promoter from (pET-29a+)(Novagen). |
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| + | We used the methods described by <em> Y. TAZUKE et al.</em> (1992) to assay sulfatase activity. Figure 5 shows that the activity of BSS was estimated at approximately 0.15 unit. | ||
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| + | <center><img src="https://static.igem.wiki/teams/4164/wiki/part-registry/bss-activity.png"with="1000" height="" width="500" height=""/></center> | ||
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| + | <p style="text-align: center!important;"><b>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).</b></p> | ||
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Latest revision as of 06:40, 12 October 2022
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:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1600
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1001
Illegal NgoMIV site found at 1052 - 1000COMPATIBLE WITH RFC[1000]