Difference between revisions of "Part:BBa K4620020"
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Arb2BURP from Coffea arabica is a split cyclase - naturally expressed separately from its peptide substrate and therefore had greater potential to cyclase various peptide substrates. We cloned Arb2BURP into MBP-pEXP expression vector. The protein was expressed in 2TY medium overnight at 18 ⁰C, purified using Ni-NTA chromatography in 8 M urea buffer, and refolded at 4 ⁰C overnight. Refolded protein was applied to 75 pg SEC column. | Arb2BURP from Coffea arabica is a split cyclase - naturally expressed separately from its peptide substrate and therefore had greater potential to cyclase various peptide substrates. We cloned Arb2BURP into MBP-pEXP expression vector. The protein was expressed in 2TY medium overnight at 18 ⁰C, purified using Ni-NTA chromatography in 8 M urea buffer, and refolded at 4 ⁰C overnight. Refolded protein was applied to 75 pg SEC column. | ||
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Latest revision as of 15:57, 12 October 2023
Arb2BURP
Arb2BURP from Coffea arabica is a split cyclase - naturally expressed separately from its peptide substrate and therefore had greater potential to cyclase various peptide substrates. We cloned Arb2BURP into MBP-pEXP expression vector. The protein was expressed in 2TY medium overnight at 18 ⁰C, purified using Ni-NTA chromatography in 8 M urea buffer, and refolded at 4 ⁰C overnight. Refolded protein was applied to 75 pg SEC column.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 211
Illegal PstI site found at 424 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 211
Illegal PstI site found at 424 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 211
Illegal PstI site found at 424 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 211
Illegal PstI site found at 424 - 1000COMPATIBLE WITH RFC[1000]