Difference between revisions of "Part:BBa K1373005"
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===oprF Verification=== | ===oprF Verification=== | ||
| − | [[Image:T--XHD-Wuhan-China--XHD-7.jpg | thumb | center | 500px |Figure | + | [[Image:T--XHD-Wuhan-China--XHD-7.jpg | thumb | center | 500px |Figure 1 Schematic illustration ]] |
Synthesis of porin gene (oprF) by direct DNA synthesis | Synthesis of porin gene (oprF) by direct DNA synthesis | ||
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In MFC operating system, Zn2+ with different concentrations (0-500 μ M) is added to the anode medium in M9 liquid culture medium, so that Zn2+ can be used in response to the regulator. The cathode solution was potassium ferricyanide (100 mM potassium ferricyanide in 50 mM phosphate buffer, pH 7.0), and voltages were recorded at 10 min intervals in an MFC biosensor using a data acquisition device. | In MFC operating system, Zn2+ with different concentrations (0-500 μ M) is added to the anode medium in M9 liquid culture medium, so that Zn2+ can be used in response to the regulator. The cathode solution was potassium ferricyanide (100 mM potassium ferricyanide in 50 mM phosphate buffer, pH 7.0), and voltages were recorded at 10 min intervals in an MFC biosensor using a data acquisition device. | ||
| − | [[Image:T--XHD-Wuhan-China--XHD-8.jpg | thumb | center | 500px |Figure | + | [[Image:T--XHD-Wuhan-China--XHD-8.jpg | thumb | center | 500px |Figure 2 Relationship between copper ion concentration and the maximum voltage of the constructed MFC biosensor ]] |
| − | [[Image:T--XHD-Wuhan-China--XHD-9.jpg | thumb | center | 500px |Figure | + | [[Image:T--XHD-Wuhan-China--XHD-9.jpg | thumb | center | 500px |Figure 3 Comparison of the maximum voltage between PLZ01-PcutR-RIBB-OPRF engineered bacteria and PLZ01-PCUTR-RIbb engineered bacteria at 500 μM copper ion concentration ]] |
The experimental results showed that oprF porin gene significantly changed the permeability of cell membrane, and its amplification effect was about 6 times. In the modified MFC sensor, there is a linear relationship between copper ion concentration and maximum voltage. | The experimental results showed that oprF porin gene significantly changed the permeability of cell membrane, and its amplification effect was about 6 times. In the modified MFC sensor, there is a linear relationship between copper ion concentration and maximum voltage. | ||
Latest revision as of 09:19, 12 October 2022
Added by XHD-Wuhan-China
oprF Verification
Synthesis of porin gene (oprF) by direct DNA synthesis
The recombinant plasmid pLZ01-pcutR-ribB-oprF was transferred to Escherichia coli BL21 for expression.
A two-chamber MFC reactor with a working volume of 240 mL was set up, and the electrodes were pretreated before use. Carbon felt with an area of 16cm2 was used as anode and cathode. These electrodes are connected by titanium wires to an external resistor of 1000 Ω.
In MFC operating system, Zn2+ with different concentrations (0-500 μ M) is added to the anode medium in M9 liquid culture medium, so that Zn2+ can be used in response to the regulator. The cathode solution was potassium ferricyanide (100 mM potassium ferricyanide in 50 mM phosphate buffer, pH 7.0), and voltages were recorded at 10 min intervals in an MFC biosensor using a data acquisition device.
The experimental results showed that oprF porin gene significantly changed the permeability of cell membrane, and its amplification effect was about 6 times. In the modified MFC sensor, there is a linear relationship between copper ion concentration and maximum voltage.
oprF gene coding porin from Pseudomonas Aeruginosa PAO1
coding sequence of oprF
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 582
- 1000COMPATIBLE WITH RFC[1000]