Difference between revisions of "Part:BBa K3185004"

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(→‎Usage and Biology)
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==Usage and Biology==
 
==Usage and Biology==
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BaCBM2 is a Carbonhydrate-binding Module family 2(CBM2) derived from Bacillus anthracis[1].CBMs are protein domains mainly found in carbohydrate-active enzymes.
+
BaCBM2 is a Carbonhydrate-binding Module family 2(CBM2) derived from Bacillus anthracis.CBMs are protein domains mainly found in carbohydrate-active enzymes.In an article[1], BaCBM2 is proved to have the highest binding ability to PET film, so we
 
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<br>
 
<br>
 
<br>
 
As our project involves connecting different proteins, we decided to use the SpyTag/SpyCatcher system and have inserted the SpyCatcher at the N-terminal of the BaCBM2.
 
As our project involves connecting different proteins, we decided to use the SpyTag/SpyCatcher system and have inserted the SpyCatcher at the N-terminal of the BaCBM2.
 +
 +
His
 +
MYC
 +
 +
TEV
  
 
We insert this part in pET-11a plasmid between BamHI and NdeI restriction sites.
 
We insert this part in pET-11a plasmid between BamHI and NdeI restriction sites.

Revision as of 11:14, 8 October 2019


SPYCatcher -> BaCBM2

Usage and Biology

BaCBM2 is a Carbonhydrate-binding Module family 2(CBM2) derived from Bacillus anthracis.CBMs are protein domains mainly found in carbohydrate-active enzymes.In an article[1], BaCBM2 is proved to have the highest binding ability to PET film, so we

As our project involves connecting different proteins, we decided to use the SpyTag/SpyCatcher system and have inserted the SpyCatcher at the N-terminal of the BaCBM2.

His MYC

TEV

We insert this part in pET-11a plasmid between BamHI and NdeI restriction sites.


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]

Expression

  • Cells were grown in 200ml LB media (100μg/ml Ampicillin) at 37oC shaking at 140 rpm to an OD600 of 0.5, verifying via a spectrophotometer.
  • Protein was expressed in 0.1mM IPTG for 2hours.

SDS-PAGE