Part:BBa_K2107000:Design
Platelet Derived Growth Factor-B Isoform 1 Serine Modified
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
When designing this Biobrick we were concerned with the ability of E. coli to correctly fold PDGF-B and form the PDGF-BB dimer because it contains four disulphide bonds. Initially, we decided to include a periplasm localization signal (DsbA) with the part, which cotranslationally translocates PDGF-B into the periplasm. This is important because the periplasm is the most optimized region of the cell for disulphide bond formation, and therefore correct folding. In addition to this we also removed two cysteine residues and replaced them with serines. By doing this we removed one of the disulphide bonds that make up PDGF-BB. Previous research has shown that this method increases the folding efficiency of PDGF-BB while only slightly increasing susceptibility to extreme pH environments.
Source
GAATTCGCGGCCGCTTCTAGAGTAGAGAAAGAGGGGACAAACTAGTATGAAAAAGATTTGGCTGGCGCTGGCTGGTTTAGTTTTAGCGTTTAGCGCATCGGCGAACCGTTGCTGGGCGCTGTTCCTGTCTCTGTGCTGCTACCTGCGTCTGGTTTCTGCGGAAGGTGACCCGATCCCGGAAGAACTGTACGAAATGCTGTCTGACCACTCTATCCGTTCTTTCGACGACCTGCAACGTCTGCTGCACGGTGATCCGGGTGAAGAAGACGGTGCGGAACTGGACCTGAACATGACCCGTTCTCACTCTGGTGGTGAACTGGAATCTCTGGCGCGTGGTCGTCGTTCTCTGGGTTCTCTGACCATCGCGGAACCGGCGATGATCGCGGAATGCAAAACCCGTACCGAAGTTTTCGAAATCTCTCGTCGTCTGATCGACCGTACCAACGCGAACTTCCTGGTTTGGCCGCCGTCTGTTGAAGTTCAGCGTTGCTCTGGTTCTTGCAACAACCGTAACGTTCAGTGCCGTCCGACCCAGGTTCAACTGCGTCCGGTTCAGGTTCGTAAAATCGAAATCGTTCGTAAAAAACCGATCTTCAAAAAAGCGACCGTTACCCTGGAAGACCACCTGGCGTGCAAATGCGAAACCGTTGCGGCGGCGCGTCCGGTTACCCGTTCTCCGGGTGGTTCTCAGGAACAGCGTGCGAAAACCCCGCAGACCCGTGTTACCATCCGTACCGTTCGTGTTCGTCGTCCGCCGAAAGGTAAACACCGTAAATTCAAACACACCCACGACAAAACCGCGCTGAAAGAAACCCTGGGTGCGCATCATCACCATCACCACTAATGATACTAGTAGCGGCCGCTGCAG