We constructed three composite parts and all of their function have been validated:Results
|K2354010||Composite||monobody-GBP HS||Bind to ER alpha when ER alpha is bind with EDCs and it can bind on the gold surface.||Chan, Wei-Jen||797|
|K2354011||Composite||INP-ER-alpha HS||Expression of ER-alpha on the membrane of E. coli and ER-alpha can bind to EDCs||Chan, Wei-Jen||2199|
|K2354012||Composite||HRP-his tag||Degrade Estrogen Disrupt Chemicals||Chan, Wei-Jen||1217|
Monobody was fused with gold binding protein and linked with 6×his tag. This fused protein can bind to the gold surface by the function of the gold-binding protein, and monobody can bind to ER-alpha/EDCs complex.
ER-alpha was fused with ice nucleating protein. This fused protein can be anchored to the membrane of E. coli via the function of ice nucleating protein and ER-alpha can bind to EDCs for detection.
Horseradish peroxidase is a kind of peroxidase which uses heme as it's cofactor, and it can degrade EDCs to less toxic compounds. Moreover, we linked fused protein with 6×his tag for purification.