Introduction
Part Collection
1. Part Collection One (Engineered Biofilm Subunit CsgA with SpyCatcher and HisTag)
a) In order to realize covalent link of two proteins, we make use of SpyTag and SpyCatcher system (see our Linkage System on Biofilm Page). The SpyTag is fused to HydA of the hydrogenase system, thus a SpyCatcher on CsgA would enable Spy-tagged Hydrogenase to be covalently attached to CsgA In the meantime, to endow biofilm CsgA protein with additional functionalities (such as specific binding to QDs or Nanorods in our case), we append one or two HisTag at the N- or/and C-terminus of the CsgA-spycatcher protein. This lead to two new constructs, HisTag-CsgA-SpyCatcher-HisTag and HisTag-CsgA-SpyCatcher (the former being sequence confirmed and submitted).
b) To determine if expression of the two proteins is successful, we constructed mCherry-SpyTag to test the activity of the SpyCatcher on the CsgA. mCherry-SpyTag is thus submitted as BBa_K2132003, sequence confirmed.
![](https://static.igem.org/mediawiki/2015/9/9e/Heidelberg_media_pictures_aachen.png)