Waiting for Loading...
First of all, you should know our number of engineering strains, and then we will use this number to explain.
| Number of engineering strains | |
|---|---|
| BMCP-B1 | E. coli K-12 MG1655 |
| BMCP-B2 | E. coli MG1655 ΔompR::kn-ccdB |
| BMCP-B3 | E. coli MG1655-ompR234 |
| BMCP-B4 | E. coli MG1655-ompR234 ΔcsgA::kn-ccdB |
| BMCP-B5 | E. coli BL21 |
-NEU-China team established
-Learn the basics of iGEM
-NEU-China team experimental group was formally established
-Establish the general direction and find relevant literature
-Complete the project: BMCP (Biofilm for metal corrosion prevention)
-The experimental group entered the laboratory in batches to participate in basic experimental training
-The experimental group was divided into four groups, and the experimental part they were responsible for was trained pertinently
-Refer to literature and optimize experimental design
-Officially start the experiment
-Electroconversion of pSIM6 vector into BMCP-B1
-Amplification of ompR-Kn-ccdB gene fragment from CR201 strain
-Construction of BMCP-B2 strain (first homologous replacement: Kn-pBAD-ccdB replacing ompR gene)
-Amplification of ompR234 gene fragment from gene synthesis fragment
-Construction of BMCP-B3 strain (second homologous replacement: ompR234 replacing Kn-pBAD-ccdB gene)
-Construction of BMCP-B4 strain (Kn-pBAD-ccdB replacing csgA gene)
-Verification of ccdB suicide switch
-PCR linearization vector pCA24N
-Amplification of mfp5 gene fragment from vector pET28a-mfp5-spycatcher
-Amplification of csgA fragment from BMCP-B1
-Construct the recombinant vector pCA24N-csgA-mfp5 by Gibson assembly
-Verification of pCA24N-csgA-mfp5 by double digestion(by Hind III，Xho I)
-Electroconversion of pCA24N-csgA-mfp5 to BMCP-B4
-BMCP-B4 pCA24N-csgA-mfp5 biofilm forming ability test: Crystal violet test, Congo red test
-PCR linearization vector pCA24N
-The csgA-spyTag gene fragment was obtained by amplifying csgA from BMCP-B1 and introducing spyTag by primers
-Construct the recombinant vector pCA24N-csgA-spyTag by Gibson assembly
-Verification of pCA24N-csgA-spyTag by double digestion(by Hind III, Xho I)
-Electroconversion of pCA24N-csgA-spyTag into BMCP-B4
-Electroconversion of pET28a-mfp5-spycatcher into BMCP-B5
-BMCP-B4 Biofilm Formation Tests: Crystal Violet Test, Congo Red Test
-Electrochemical test of corrosion of functional biofilm in vivo
-OCP curve of corrosion electrochemical test
-LPR curve of corrosion electrochemical test
-Polarization curve of corrosion electrochemical test
-EIS curve of corrosion electrochemical test
-EIS curve after coating on electrode for 1 day, left: Nyquist plot, right: Bode plot
-EIS curve after coating on electrode for 7 days, left: Nyquist plot, right: Bode plot
-Nyquist plot of four groups of materials after one day, three days, five days and seven days
-Bode plot of four groups of materials after one day, three days, five days and seven days
-Biofilm adhesion test in vivo (80rpm shaker)
-Characterization of biomineralization morphology of biofilm by SEM
-Observation on anticorrosion effect of mineralized biofilm
