Difference between revisions of "Part:BBa K523016"
 
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This is ''C. fimi'' exoglucanase under the control of the Lac promoter. This was made by Edinburgh 2008 but vector-swapped into pSB1C3 and entered into the Registry by Edinburgh 2011.
 
This is ''C. fimi'' exoglucanase under the control of the Lac promoter. This was made by Edinburgh 2008 but vector-swapped into pSB1C3 and entered into the Registry by Edinburgh 2011.
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===Characterization===
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Edinburgh 2011 conducted two assays, comparing the activity of this part to a &beta;-glucosidase (''E. coli'' bglX, <partinfo>BBa_K523014</partinfo>, also under the control of the lac promoter) on two different substrates:
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* 4-methylumbelliferyl &beta;- D- glucuronide '''(MUG, left photo)'''. This substrate is a cellobiose analog.
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* 4-methylumbelliferyl &beta;- D- cellobioside '''(MUC, right photo)'''. This substrate is larger and is more like a cellulose analog.
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Both substrates produce a fluorescent product when cleaved. Our plates below show the results of placing cell lysate and cell debris on an MUG plate and an MUC plate. Present on both plates are:
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* Left side of plate: lysate/debris from JM109 expressing ''bglX'', <partinfo>BBa_K523014</partinfo>
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* Right side of plate: lysate/debris from JM109 expressing ''cex'', <partinfo>BBa_K523016</partinfo>
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* Bottom of plate: lysate/debris from JM109 cells
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<center>
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{| style="margin-top: 1em; margin-bottom: 1em;"
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|-
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|[[Image:BglX-MuG.jpg|300px]]
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|&nbsp; &nbsp;
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|[[Image:Cex-MuC.jpg|300px]]
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|-
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|width="300px" valign="top"|'''MUG assay.''' ''bglX'' on left, ''cex'' on right.
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|&nbsp; &nbsp;
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|width="300px" valign="top"|'''MUC assay.''' ''bglX'' on left, ''cex'' on right.
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|}
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</center>
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As can be seen, ''cex'' is much better at degrading MUC.
  
  

Latest revision as of 20:24, 7 October 2011

Plac + RBS + cex (exoglucanase)

This is C. fimi exoglucanase under the control of the Lac promoter. This was made by Edinburgh 2008 but vector-swapped into pSB1C3 and entered into the Registry by Edinburgh 2011.

Characterization

Edinburgh 2011 conducted two assays, comparing the activity of this part to a β-glucosidase (E. coli bglX, BBa_K523014, also under the control of the lac promoter) on two different substrates:

  • 4-methylumbelliferyl β- D- glucuronide (MUG, left photo). This substrate is a cellobiose analog.
  • 4-methylumbelliferyl β- D- cellobioside (MUC, right photo). This substrate is larger and is more like a cellulose analog.

Both substrates produce a fluorescent product when cleaved. Our plates below show the results of placing cell lysate and cell debris on an MUG plate and an MUC plate. Present on both plates are:

  • Left side of plate: lysate/debris from JM109 expressing bglX, BBa_K523014
  • Right side of plate: lysate/debris from JM109 expressing cex, BBa_K523016
  • Bottom of plate: lysate/debris from JM109 cells
BglX-MuG.jpg     Cex-MuC.jpg
MUG assay. bglX on left, cex on right.     MUC assay. bglX on left, cex on right.

As can be seen, cex is much better at degrading MUC.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1148
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 781
    Illegal NgoMIV site found at 1154
    Illegal NgoMIV site found at 1656
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1201
    Illegal SapI.rc site found at 1284