Part:BBa_K3185004

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.According to an article[1], BaCBM2 is proved to have the highest binding ability to PET film, so we use this protein as

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 37^oC shaking at 140 rpm to an OD600 of 0.5, verifying via a spectrophotometer.

• Protein was expressed in 0.1mM IPTG for 2hours.