BBa_K243002
1
DsbA
DsbA signal sequence (enables periplasm export)
2009-10-07T11:00:00Z
2015-05-08T01:11:37Z
Sequence of the DsbA was copied out from E.coli B genom
Synthesized oligos with signal sequence of DsbA by sigma.
The signal sequence of DsbA linked to an cds of e.g.a protein, applies the exportation of the protein to the periplasm. The accumulation of the expressed protein in the periplasm, can be used for the purification of the protein.
false
false
_352_
0
4732
9
It's complicated
false
the signal sequence had to be located at the N-terminus of the (fusion)protein.
false
Freiburg Bioware09
annotation2041875
1
Dsba signal sequence
range2041875
1
1
54
BBa_K2107010
1
BBa_K2107010
Platelet Derived Growth Factor-B Isoform 1 (Serine Modified)
2016-10-18T11:00:00Z
2016-10-20T09:27:42Z
GAATTCGCGGCCGCTTCTAGAGTAGAGAAAGAGGGGACAAACTAGTATGAAAAAGATTTGGCTGGCGCTGGCTGGTTTAGTTTTAGCGTTTAGCGCATCGGCGAACCGTTGCTGGGCGCTGTTCCTGTCTCTGTGCTGCTACCTGCGTCTGGTTTCTGCGGAAGGTGACCCGATCCCGGAAGAACTGTACGAAATGCTGTCTGACCACTCTATCCGTTCTTTCGACGACCTGCAACGTCTGCTGCACGGTGATCCGGGTGAAGAAGACGGTGCGGAACTGGACCTGAACATGACCCGTTCTCACTCTGGTGGTGAACTGGAATCTCTGGCGCGTGGTCGTCGTTCTCTGGGTTCTCTGACCATCGCGGAACCGGCGATGATCGCGGAATGCAAAACCCGTACCGAAGTTTTCGAAATCTCTCGTCGTCTGATCGACCGTACCAACGCGAACTTCCTGGTTTGGCCGCCGTCTGTTGAAGTTCAGCGTTGCTCTGGTTCTTGCAACAACCGTAACGTTCAGTGCCGTCCGACCCAGGTTCAACTGCGTCCGGTTCAGGTTCGTAAAATCGAAATCGTTCGTAAAAAACCGATCTTCAAAAAAGCGACCGTTACCCTGGAAGACCACCTGGCGTGCAAATGCGAAACCGTTGCGGCGGCGCGTCCGGTTACCCGTTCTCCGGGTGGTTCTCAGGAACAGCGTGCGAAAACCCCGCAGACCCGTGTTACCATCCGTACCGTTCGTGTTCGTCGTCCGCCGAAAGGTAAACACCGTAAATTCAAACACACCCACGACAAAACCGCGCTGAAAGAAACCCTGGGTGCGCATCATCACCATCACCACTAATGATACTAGTAGCGGCCGCTGCAG
This Biobrick codes for Platelet Derived Growth Factor-Beta Isoform 1, it is a growth factor that has been shown to modulate cell proliferation in angiogenesis. Additionally, PDGF-B is preceded by the periplasmic localization signal DsbA. When imaging this PDGF-B in western blot it has been shown to appear between 28 and 37Kda. To help with protein purification this part also includes a His tag at the c-terminus.
true
false
_2575_
30500
30500
9
false
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.
This part should be cloned into a low to medium copy plasmid and transformed into cells designed for recombinant protein expression such as Origami E. coli.
false
iGEM Raiders
component2524193
1
BBa_K243002
annotation2524193
1
BBa_K243002
range2524193
1
1
54
BBa_K2107010_sequence
1
aaaaagatttggctggcgctggctggtttagttttagcgtttagcgcatcggcg
BBa_K243002_sequence
1
aaaaagatttggctggcgctggctggtttagttttagcgtttagcgcatcggcg
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z