Difference between revisions of "Part:BBa K2985009"
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<p>SDS-page detection qualitative analysis
 
<p>SDS-page detection qualitative analysis
 
<h4>Methods and results</h4>
 
<h4>Methods and results</h4>
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<p>Take 1 mL fermented liquid, in the 12000r∙min-1, 1 min the centrifugal, the fermented supernatant as samples to identify the Protein SDS-page electrophoresis of pullulan enzyme expression, using SDS-PAGE of concentrated gum concentration was 5%, the concentration of separation gel was 9%, take samples of 30 mu, 4 x L SDS-page Protein Loading Buffer 10 mu L and blending boil after 5 min, 5000 r, 2 min min-1 centrifugal, samples from 15 mu L processed Protein to join the fibrin glue lane, 5 μl high molecular weight protein Marker was added.<br>
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Take 1 mL fermented liquid, in the 12000r∙min-1, 1 min the centrifugal, the fermented supernatant as samples to identify the Protein SDS-page electrophoresis of pullulan enzyme expression, using SDS-PAGE of concentrated gum concentration was 5%, the concentration of separation gel was 9%, take samples of 30 mu, 4 x L SDS-page Protein Loading Buffer 10 mu L and blending boil after 5 min, 5000 r, 2 min min-1 centrifugal, samples from 15 mu L processed Protein to join the fibrin glue lane, 5 μl high molecular weight protein Marker was added.<br>
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<p>
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<img src="https://parts.igem.org/File:Figure_1-pul.jpeg" style="width:40%">
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Revision as of 00:48, 20 October 2019


Pul-> Amylopectin 6-glucan hydrolase

Pul gene encodes a protein, pullulanase, which can specifically hydrolyze the alpha-1,6 glycoside bonds in amylopectin branching points by endonucleation, thus cutting down the whole branching structure to form amylose amylopectin debranching enzymes. We use these enzymes as a tool for our characterization of promoters.

SDS-page detection qualitative analysis

Methods and results

Take 1 mL fermented liquid, in the 12000r∙min-1, 1 min the centrifugal, the fermented supernatant as samples to identify the Protein SDS-page electrophoresis of pullulan enzyme expression, using SDS-PAGE of concentrated gum concentration was 5%, the concentration of separation gel was 9%, take samples of 30 mu, 4 x L SDS-page Protein Loading Buffer 10 mu L and blending boil after 5 min, 5000 r, 2 min min-1 centrifugal, samples from 15 mu L processed Protein to join the fibrin glue lane, 5 μl high molecular weight protein Marker was added.
<p> <img src="https://parts.igem.org/File:Figure_1-pul.jpeg" style="width:40%">



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]