Part:BBa_K771008
Membrane Anchor 5 without VVD:ssDsbA-LGT-GBD Ligand
Membrane anchor 5 without VVD, which consists of GBD ligand(BBa_K771106) and membrane protein Lgt(BBa_K771102). Membrane Anchor 5 (with and without VVD) is proved to constitutively dimerize with Membrane Anchor [BBa_K771004].
Overview: Membrane Scaffold System (without signal induction)
Demonstration of the Membrane Scaffold device (without signal induction). Membrane Anchor 2 (without MS2), 3, 4, 5(without VVD) consitutively aggregate
together.
Backgroud
Constitutive Aggregation
Membrane Anchor 4 (BBa_K771003)and Membrane Anchor 5 (with and without VVD) constitutively
aggregate through GBD domain(BBa_K771105) and GBD ligand
(BBa_K771106).
Characterization
We proved the constitutive interaction between membrane anchor 4 and 5 by fusing split EGFP to membrane anchor 4 and membrane anchor 5 without VVD.
Membrane Anchor 4 and Membrane Anchor 5 without VVD
We fused split EGFP 1 and 2 to Membrane Anchor 4 and Membrane Anchor 5 without VVD to test whether Membrane Anchor 4 and Membrane Anchor 5 without VVD could constitutively aggregate. Proteins in control group are not expected to aggregate like in experimental group.We coexpressed proteins in experimental group and control group respectively in E.coli. After induction for 6 hours, bacteria samples were taken for fluorescence observationThe result shows that Membrane Anchor 4 and Membrane Anchor 5 without VVD could constitutively aggregate through GBD domain and ligand.
Design
Membrane Anchor 5 could constitutively aggregate with Membrane Anchor 4.
Application
We recruired fatty acid biosynthetic pathway involving TesA(BBa_K771301), FabG(BBa_K771302)) , FabI(BBa_K771303), FabZ(BBa_K771304) to put our membrane scaffold system into practice. Fatty Acid productivity is enhanced by 24 fold.
We constructed TesA with Membrane Anchor 2 (without MS2) (BBa_K771305),FabG with Membrane
Anchor 3,(BBa_K771306), FabZ with Membrane Anchor 4(BBa_K771307), FabI with Membrane Anchor 4(without VVD ) (BBa_K771308). Click into each part for more infomation.
Related Biobrick
<br\>Membrabe Anchor 2 without MS2([BBa_K771007]) <br\>Membrabe Anchor 3([BBa_K771003]) <br\>Membrabe Anchor 4([BBa_K771004])
Reference
Dueber, J. E., G. C. Wu, et al. (2009). "Synthetic protein scaffolds provide modular control over metabolic flux." Nature biotechnology 27(8): 753-759.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 883
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 504
n/a | Membrane Anchor 5 without VVD:ssDsbA-LGT-GBD Ligand |