Difference between revisions of "Team:HUST-China/Experiments"

2. Add Flag Sequence into parts

The constructed plasmid was transformed into E.coli Top 10.

3. Add 6×His Sequence into parts

The constructed plasmid was transformed into E.coli Top 10.

4. pck360 Plasmid Conjugation

We constructed following vectors carrying our parts and transformed these vectors into E.coli Top 10 to transformed into cynobacteria.

Figure 6. modified circuit of ldhD-lldP.

Figure 7. modified circuit of ldhDC-lldP.

Figure 8. modified circuit of ldhDnARSdR-lldP.

Figure 9. modified circuit of ldhDARSdR-lldP.

5. Detection of yellow fluorescent protein

Since plasmid pck306 has yellow fluorescent protein sequence, we cultivated the engineering cynobacteria induced by rhamnose for 24h. And put them under the fluorescence microscope at the wave of 488nm excitation light

2. Transformation into E.coli BL21

The constructed plasmid was transformed into E.coli BL21.

3. Transformation into Rhodopseudomonas palustris DSM5859

The constructed plasmid was transformed into Rhodopseudomonas palustris DSM5859.

4. Verification the expression on mRNA level

We did RT-qPCR to test the relative expression levels of each genes.

5. Detection of Lactate

We detect lactate by Lactic Acid assay kit.

2. Transformation into E.coli WM3064

The constructed plasmid was transformed into a 2,6-diaminopimelic acid deficiency type E.coli WM3064.

3. Conjugation between E.coli and Shewanella oneidensis MR-1

We did conjugation after the transformation to WM3064 to introduce the plasmid into Shewanella oneidensis MR-1.

4. Verification the expression on mRNA level

We did RT-qPCR to test the relative expression levels of each genes.

5. Electrogenesis detection

We cultivated the engineering bacteria and put them into the electrogenesis device to test the output votage. We also did the measurement of electrode-attached biomass.

Co-culture and electricity

1. Contrast the symbiotic effect of wild-type strains

MFC anode: Synechocystis PCC6803 (wild type) + Shewanella

MFC anode: Rhodopseudomonas palustris (wild type) + Shewanella

2. Functional verification of engineered Synechococcus PCC6803

MFC anode: Synechocystis PCC6803 (wild type) + Shewanella

MFC anode: Synechocystis PCC6803 (engineered type) + Shewanella

3. Functional verification of engineered Rhodopseudomonas palustris

MFC anode: Rhodopseudomonas palustris (wild type) + Shewanella

MFC anode: Rhodopseudomonas palustris (engineered type) + Shewanella

4. Study the effects of oxygen on Co-culture power systems.

MFC anode: anaerobic

MFC anode: aerobic

5. Demonstrate that carbon cloth has higher electricity production efficiency than carbon rod.

MFC anode: carbon cloth

MFC anode: carbon rod