Difference between revisions of "Part:BBa K1373005"

 
Line 1: Line 1:
 +
==Added by XHD-Wuhan-China==
 +
 +
===oprF Verification===
 +
 +
[[Image:T--XHD-Wuhan-China--XHD-7.jpg | thumb | center | 500px |Figure 7 Schematic illustration ]]
 +
 +
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.
 +
 +
[[Image:T--XHD-Wuhan-China--XHD-8.jpg | thumb | center | 500px |Figure 8 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 9 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.
 +
 +
 +
  
 
__NOTOC__
 
__NOTOC__

Revision as of 09:15, 12 October 2022

Added by XHD-Wuhan-China

oprF Verification

Figure 7 Schematic illustration

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.

Figure 8 Relationship between copper ion concentration and the maximum voltage of the constructed MFC biosensor
Figure 9 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.



oprF gene coding porin from Pseudomonas Aeruginosa PAO1

coding sequence of oprF

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 582
  • 1000
    COMPATIBLE WITH RFC[1000]