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Revision as of 10:59, 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.