Difference between revisions of "Part:BBa K4385025"
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<partinfo>BBa_K4385025 short</partinfo>
 
<partinfo>BBa_K4385025 short</partinfo>
  
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This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption protein. The PbrR without absorbing lead ions was connected and formed a complex under TurbolD's traction. With the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. So we can detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.
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This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption protein.  
  
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===Usage and Biology===
 
===Usage and Biology===
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The PbrR without absorbing lead ions was connected and formed a complex under TurbolD's traction. With the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. So we can detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.
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===Characterization===
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Our surface display system can express the Lpp-OmpA-pbrR protein in the outer membrane.So we use the turboid-pbrR to test the pbrR function.With the expression of Lpp-OmpA-pbrR, our bacteria have the ability to adsorb the Pb2+.
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[[Image: TP.png|center|frame|100px|<b>Figure 1. TurboID-PbrR</b>]]<br><br>
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Our purpose is to detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.And with the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed.
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[[Image: TP2.png|center|frame|100px|<b>Figure 2. The different concentration of the Pb2+ in the culture medium </b> (A)0mM;(B)1μM;(C)10μM;(D) 1mM.DAPI channel was used the concentration of the bacteria,the Cy3 channel was used to verify the quantity of the pbrR protein which is not combined with the Pb2+.The result showed that the number of the monomer pbrR protein decreased when there is a higher concentration of the Pb2+.This provided us with the possibility to evaluate the function of our surface display system. ]]<br><br>
  
 
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Revision as of 07:32, 12 October 2022


PbrR location detection

This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption protein.

Usage and Biology

The PbrR without absorbing lead ions was connected and formed a complex under TurbolD's traction. With the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. So we can detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.

Characterization

Our surface display system can express the Lpp-OmpA-pbrR protein in the outer membrane.So we use the turboid-pbrR to test the pbrR function.With the expression of Lpp-OmpA-pbrR, our bacteria have the ability to adsorb the Pb2+.

Figure 1. TurboID-PbrR


Our purpose is to detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.And with the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed.

Figure 2. The different concentration of the Pb2+ in the culture medium (A)0mM;(B)1μM;(C)10μM;(D) 1mM.DAPI channel was used the concentration of the bacteria,the Cy3 channel was used to verify the quantity of the pbrR protein which is not combined with the Pb2+.The result showed that the number of the monomer pbrR protein decreased when there is a higher concentration of the Pb2+.This provided us with the possibility to evaluate the function of our surface display system.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1486
    Illegal PstI site found at 1428
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 1486
    Illegal PstI site found at 1428
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1486
    Illegal PstI site found at 1428
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1486
    Illegal PstI site found at 1428
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1219