Difference between revisions of "Part:BBa K523014"
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We conducted two assays, comparing the activity of this part ''(bglX)'' with that of the exoglucanase cex (<partinfo>BBa_K118022</partinfo>) on two different substrates: | We conducted two assays, comparing the activity of this part ''(bglX)'' with that of the exoglucanase cex (<partinfo>BBa_K118022</partinfo>) on two different substrates: | ||
− | * 4-methylumbelliferyl-beta-D-glucuronide (MUG). This substrate mimics cellobiose but produces a fluorescent product when cleaved. | + | * 4-methylumbelliferyl-beta-D-glucuronide '''(MUG, left photo)'''. This substrate mimics cellobiose but produces a fluorescent product when cleaved. |
− | * Something else. | + | * Something else '''(MUC, right photo)'''. |
Our plates below shows the results of placing cell lysate and cell debris on an MUG plate. Present on both plates are: | Our plates below shows the results of placing cell lysate and cell debris on an MUG plate. Present on both plates are: |
Revision as of 13:15, 13 September 2011
Plac + LacZ + bglX
The E. coli periplasmic β-glucosidase gene bglX under the control of the lac promoter. The native ribosome binding site is present.
Usage and Biology
The BglX β-glucosidase is expected to be localised to the periplasm. It ought to be capable of breaking down cellobiose.
Experiment
We conducted two assays, comparing the activity of this part (bglX) with that of the exoglucanase cex (BBa_K118022) on two different substrates:
- 4-methylumbelliferyl-beta-D-glucuronide (MUG, left photo). This substrate mimics cellobiose but produces a fluorescent product when cleaved.
- Something else (MUC, right photo).
Our plates below shows the results of placing cell lysate and cell debris on an MUG plate. Present on both plates are:
- Left side of plate: JM109 expressing this part, K523014
- Right side of plate: JM109 expressing exoglucanase
- Bottom of plate: JM109 cells
Foo. | Bar. |
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 607
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2266
Illegal AgeI site found at 2488
Illegal AgeI site found at 2677 - 1000COMPATIBLE WITH RFC[1000]