Difference between revisions of "Part:BBa K3185004"
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<partinfo>BBa_K3185004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3185004 SequenceAndFeatures</partinfo>
 
==Cultivation and Expression==
 
==Cultivation and Expression==
 +
Expression:
 +
<ul>
 +
<li>Cells were grown in 200ml LB media (100μg/ml Ampicillin) at 37<sup>o</sup>C shaking at 140 rpm to an OD600 of 0.5, verifying via a spectrophotometer.
 +
</ul>
 +
<ul>
 +
<li>Protein was expressed in 0.1mM IPTG for 2hours.
 +
</ul>
  
  

Revision as of 10:58, 8 October 2019


SPYCatcher -> BaCBM2

Usage and Biology

BaCBM2 is a Carbonhydrate-binding Module family 2(CBM2) derived from Bacillus anthracis[1]. CBMs are protein domains mainly found in carbohydrate-active enzymes.

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.

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]

Cultivation and Expression

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.