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, left photo)'''. This substrate mimics cellobiose but produces a fluorescent product when cleaved.
+
* 4-methylumbelliferyl-beta-D-glucuronide '''(MUG, left photo)'''. This substrate mimics cellobiose.
 +
* Something else '''(MUC, right photo)'''. This substrate mimics cellulose.
  
−
* Something else '''(MUC, right photo)'''.
+
Both substrates produce a fluorescent product when cleaved. 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:
+
  
 
* Left side of plate: JM109 expressing this part, K523014
 
* Left side of plate: JM109 expressing this part, K523014
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|}
 
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</center>
 
</center>
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As can be seen, ''bglX'' is capable of degrading MUG (the cellobiose mimic) while exoglucanase is not. By contrast, ''bglX'' cannot degrade MUC (the cellulose mimic) while exoglucanase can.
  
 
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Revision as of 13:17, 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.
  • Something else (MUC, right photo). This substrate mimics cellulose.

Both substrates produce a fluorescent product when cleaved. 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
BglX-MuG.jpg     Cex-MuC.jpg
Foo.     Bar.

As can be seen, bglX is capable of degrading MUG (the cellobiose mimic) while exoglucanase is not. By contrast, bglX cannot degrade MUC (the cellulose mimic) while exoglucanase can.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 607
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2266
    Illegal AgeI site found at 2488
    Illegal AgeI site found at 2677
  • 1000
    COMPATIBLE WITH RFC[1000]