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Revision as of 17:24, 17 October 2018
the gene of csgA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
CsgA is the monomer of curli, which is an important component of excellular matrix of Enterobacteriaceae. Curli fibers are involved in adhesion to surfaces, cell aggregation, and biofilm formation, they are also used for studies of amyloid fiber formation in human diseases, such as Alzheimer’s, Huntington’s, and prion diseases.
As an important component of excellular matrix and biofilms, modified csgA provides us many potentials to remould biofilm into a brand-new functional biology matrix. We can tack many fancy characteristics to biofilm and modified bacteria. Team ZJU-CHINA 2018 made use of the design of cell-surface display and modified the csgA protein by fusing spytag to the N-terminal, making it a dorable platform to form enzyme complex and increasd the electrical conductivity because of the histag in the C-terminal.Actually, the cell-surface display has many potential applications, including biodegradation,live vaccine development, peptide library screening, bioconversion using whole cell biocatalyst and bioadsorption.
As the excellular matrix of bacteria, the curli can also interact with intracellular reactions, and function as a detector of environment or messenger between extracellular and intracellular. For example, the modified curli can be used as a membrane-based bisensor for pathogen, heavy metal and other environment pollutants.
Approach and Result
SDS-PAGE results:
To verify the over-expression of curli, we did Cango Red(CR) deletion assay.The CR can binds to amyloid proteins and thus can be pulled down to the pellet while the absorbance of supernatant will reduced.The results are as below.
Quantitative results:
We measured the absorbance of supernatant at 480nm for CR and 600nm for cell concentration.The comparison of CgsA knockout Defect bacteria, wild type BL21(DE3) our recombinant csgA-expressing bacteria shows the overexpression of our recombinant strain.